Biophotonic materials and uses thereof

ABSTRACT

The present disclosure provides biophotonic materials and methods useful in phototherapy. In particular, the biophotonic materials of the present disclosure include a cohesive matrix, and at least one chromophore, wherein the at least one chromophore can absorb and emit light from within the biophotonic material. The biophotonic materials and the methods of the present disclosure are useful for promoting wound healing and skin rejuvenation, as well as treating acne and various skin disorders.

CROSS-RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application Ser.No. 61/786,197, filed Mar. 14, 2013, and U.S. Provisional ApplicationSer. No. 61/873,747, filed Sep. 4, 2013, the entire contents of each ofwhich are incorporated herein by reference.

BACKGROUND OF THE DISCLOSURE

Phototherapy has recently been recognized as having wide range ofapplications in both the medical and cosmetic fields including use insurgery, therapy and diagnostics. For example, phototherapy has beenused to treat cancers and tumors with lessened invasiveness, todisinfect target sites as an antimicrobial treatment, to promote woundhealing, and for facial skin rejuvenation.

Photodynamic therapy is a type of phototherapy involving the applicationof a photosensitive agent to target tissue then exposing the targettissue to a light source after a determined period of time during whichthe photosensitizer is absorbed by the target tissue. Such regimens,however, are often associated with undesired side-effects, includingsystemic or localized toxicity to the patient or damage to non-targetedtissue. Moreover, such existing regimens often demonstrate lowtherapeutic efficacy due to, for example, the poor selectivity of thephotosensitive agents into the target tissues.

Therefore, it is an object of the present disclosure to provide new andimproved compositions and methods useful in phototherapy.

SUMMARY OF THE DISCLOSURE

The present disclosure provides topical biophotonic materials andmethods useful in phototherapy. In particular, the biophotonic materialsof the present disclosure include a cohesive matrix, and at least onechromophore, wherein the at least one chromophore can absorb and emitlight from within the biophotonic material.

In certain embodiments of any of the foregoing or following, thebiophotonic material is an elastic material. In certain embodiments, theelastic material is a peelable film. In other embodiments, thebiophotonic material is a non-elastic material. In some embodiments, thebiophotonic material is rigid.

In certain embodiments of any of the foregoing or following, thebiophotonic material has a tear and/or tensile strength greater than anadhesive strength of the biophotonic material to a surface to which itis applied.

In certain embodiments of any of the foregoing or following, thebiophotonic material is substantially translucent. In some embodiments,the biophotonic material has a translucency of at least about 40%, about50%, about 60%, about 70%, or about 80% at 460 nm.

In certain embodiments of any of the foregoing or following, thebiophotonic material has a thickness of about 0.1 mm to about 50 mm.

In certain embodiments of any of the foregoing or following, thebiophotonic material has a pre-formed configuration. In someembodiments, the pre-formed configuration is a shape and/or a sizecorresponding with a shape and/or a size of a body part to which thebiophotonic material can be applied. In some embodiments, the body partto which the material is applied is a head, scalp, forehead, nose,cheeks, ears, lip, face, neck, shoulder, arm pit, arm, elbow, hand,finger, abdomen, chest,stomach, back, sacrum, buttocks, genitals, legs,knee, feet, nails, hair, toes, or bony prominences, or combinationsthereof.

In certain embodiments of any of the foregoing or following, thematerial is a mask. In some embodiments, the mask is a face mask havingat least one opening for the eyes, nose or mouth.

In certain embodiments of any of the foregoing or following, thebiophotonic material has a pre-formed configuration and the pre-formedconfiguration is a shape and/or a size corresponding with a shape and/ora size of a light source or lamp to which the biophotonic material canbe attached.

In certain embodiments of any of the foregoing or following, thebiophotonic material can be removed without leaving substantially anyresidue on a surface to which the biophotonic material is applied.

In certain embodiments of any of the foregoing or following, thebiophotonic material the at least one chromophore included in thebiophotonic material is a fluorophore. In some embodiments, thefluorophore is a xanthene dye. In some embodiments, the chromophore isincluded in the cohesive matrix.

In certain embodiments of any of the foregoing or following, thecohesive matrix of the biophotonic material includes at least onepolymer. In some embodiments, the polymer is selected from across-linked polyacrylic polymer, a hyaluronate, a hydrated polymer, ahydrophilic polymer. In some embodiments, cohesive matrix comprisessodium hyaluronate. In sonic embodiments, sodium hyaluronate is presentin an amount of about 2% to about 8%. In some embodiments, the cohesivematrix comprises a carbomer. In some embodiments, the carbomer ispresent in an amount of about 0.1% to about 2%.

In certain embodiments of any of the foregoing or following, thecohesive matrix is in particulate form.

In some embodiments, the chromophore is included in a carrier mediumwhich can form a cohesive matrix. In some embodiments, the chromophorecan absorb and emit light within the cohesive matrix when illuminatedwith light. In some embodiments, the carrier medium is at least onepolymer or a polymer pre-cursor which can form the cohesive matrix bypolymerizing, cross-linking or drying.

The biophotonic material of any embodiments of the disclosure may beused as a mask, dressing or filler. The biophotonic material of anyembodiments of the disclosure may also be used for cosmetic or medicaltreatment of tissue. In some embodiments, the cosmetic treatment is skinrejuvenation and conditioning, and the medical treatment is woundhealing, periodontal treatment or acne treatment.

The present disclosure also provides containers comprising thebiophotonic material or precursor material according to variousembodiments of the disclosure. In some embodiments, the containercomprises a sealed chamber for holding a biophotonic material, and anoutlet in communication with the chamber for discharging the biophotonicmaterial from the container, wherein the biophotonic material comprisesat least one chromophore in a carrier medium which can form a cohesivematrix after being discharged from the sealed chamber. In someembodiments, the container is a spray can.

The present disclosure also provides kits for preparing or providing thebiophotonic material or precursor according to various embodiments ofthe disclosure. In some embodiments, the kit comprises a first containercomprising a first chromophore; and a second component comprising athickening agent, wherein the thickening agent can form a cohesivematrix when mixed with the first component.

The present disclosure also provides methods for biophotonic treatmentcomprising applying the biophotonic material of the disclosure to atarget tissue and illuminating the material with light. In someembodiments, the method is for biophotonic treatment of a skin disorderwherein the method comprises placing a biophotonic material over atarget skin tissue, wherein the biophotonic material comprises at leastone chromophore and a cohesive matrix; and illuminating said biophotonicmaterial with light having a wavelength that overlaps with an absorptionspectrum of the at least one chromophore; wherein said biophotonicmaterial emits fluorescence at a wavelength and intensity that promoteshealing of said skin disorder.

In some embodiments, the method is for biophotonic treatment of acnewherein the method comprises placing a biophotonic material over atarget skin tissue, wherein the biophotonic material comprises at leastone chromophore and a cohesive matrix; and illuminating said biophotonicmaterial with light having a wavelength that overlaps with an absorptionspectrum of the at least one chromophore; wherein said biophotonicmaterial emits fluorescence at a wavelength and intensity that treatsthe acne.

In some embodiments, the method is for promoting wound healing whereinthe method comprises placing a biophotonic material over or within awound, wherein the biophotonic material comprises at least onechromophore and a cohesive matrix; and illuminating said biophotonicmaterial with light having a wavelength that overlaps with an absorptionspectrum of the at least one chromophore; wherein said biophotonicmaterial emits fluorescence at a wavelength and intensity that promoteswound healing.

In some embodiments, the method is for promoting skin rejuvenationwherein the method comprises placing a biophotonic material over atarget skin tissue, wherein the biophotonic material comprises at leastone chromophore and a cohesive matrix; and illuminating said biophotonicmaterial with light having a wavelength that overlaps with an absorptionspectrum of the at least one chromophore; wherein said biophotonicmaterial emits fluorescence at a wavelength and intensity that promotesskin rejuvenation.

In some embodiments, the biophotonic material is removed afterillumination. In some embodiments, the material is peeled off afterillumination.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the absorption and emission spectra of donor andacceptor chromophores. The spectral overlap between the absorptionspectrum of the acceptor chromophore and the emission spectrum of thedonor chromophore is also shown.

FIG. 2 is a schematic of a Jablonski diagram that illustrates thecoupled transitions involved between a donor emission and acceptorabsorbance.

FIG. 3 is an emission fluorescence spectrum from an activatedbiophotonic material according to an embodiment of the presentdisclosure (Example 1).

FIG. 4 is an emission fluorescence spectrum from a photoactivatedbiophotonic material iraddiating fibroblasts and keratinocytes forevaluating protein regulation and gene expression (Example 5).

FIGS. 5a and 5b are emission fluorescence spectra for Eosin Y andFluorescein, respectively, and the activating light passing through thecomposition, at different concentrations of the chromophores (Example 7)

FIGS. 6a and 6b are absorbance and emission spectra, respectively, ofEosin and Fluorescein in a gel (Example 8).

FIGS. 7a and 7b are absorbance and emission spectra, respectively, ofEosin, Fluorescein and Rose Bengal in a gel (Example 9).

DETAILED DESCRIPTION

Overview

The present disclosure provides biophotonic materials and uses thereof.Biophotonic therapy using these materials would not involve substantialdirect contact of a photosensitive agent (or chromophore) with thetherapeutic target, which includes, but is not limited to, skin, mucousmembranes, wounds, hair and nails. Therefore, undesired side effectscaused by such direct contact may be reduced, minimized, or prevented.Furthermore, in certain embodiments, phototherapy using the biophotonicmaterials of the present disclosure will for instance rejuvenate theskin by, e.g., promoting collagen synthesis, promote wound healing,treat skin conditions such as acne, and treat periodontitis.

(2) Definitions

Before continuing to describe the present disclosure in further detail,it is to be understood that this disclosure is not limited to specificcompositions or process steps, as such may vary. It must be noted that,as used in this specification and the appended claims, the singular form“a”, “an” and “the” include plural referents unless the context clearlydictates otherwise.

As used herein, the term “about” in the context of a given value orrange refers to a value or range that is within 20%, preferably within10%, and more preferably within 5% of the given value or range.

It is convenient to point out here that “and/or” where used herein is tobe taken as specific disclosure of each of the two specified features orcomponents with or without the other. For example “A and/or B” is to betaken as specific disclosure of each of (i) A, (ii) B and (iii) A and B,just as if each is set out individually herein,

“Biophotonic” means the generation, manipulation, detection andapplication of photons in a biologically relevant context. In otherwords, biophotonic compositions and materials exert their physiologicaleffects primarily due to the generation and manipulation of photons.

“Biophotonic material” is a material which may be activated by light toproduce photons for biologically relevant applications. Biophotonicmaterials, as referred to herein, may be cohesive gels, semi-solids orsolids, that The biophotonic material can be in the form of, including,but not limited to, a film or an elastic or non-elastic matrix, or thelike, for uses such as a mask, a dressing or a light attachment. Thebiophotonic material can be a composite and include fibres,particulates, ribs, supporting structures, networks, non-biophotoniclayers or biophotonic layers with the same or different compositions.

“Cohesive matrix” refers to a medium which is, or which can form, aself-supporting material e.g. a material with a defined shape understeady state conditions. This may be due to internal attractive forces.The property of cohesion in a material can allow the material to behandled without tearing.

“Topical application” or “topical uses” means application to bodysurfaces, such as the skin, mucous membranes, vagina, oral cavity,internal surgical wound sites, and the like.

Terms “chromophore” and “photoactivator” are used hereininterchangeably. A chromophore means a chemical compound, when contactedby light irradiation, is capable of absorbing the light. The chromophorereadily undergoes photoexcitation and can transfer its energy to othermolecules or emit it as light (fluorescence).

“Photobleaching” or “photobleaches” means the photochemical destructionof a chromophore. A chromophore may fully or partially photobleach.

The term “actinic light” is intended to mean light energy emitted from aspecific light source (e.g. lamp, LED, or laser) and capable of beingabsorbed by matter (e.g. the chromophore or photoactivator). In apreferred embodiment, the actinic light is visible light.

A “peel-off” or “peelable” film or matrix is one that can bemechanically removed, such as by hand, after application. It can beremoved as a single piece, or as a small number of large pieces.

“Skin rejuvenation” means a process of reducing, diminishing, retardingor reversing one or more signs of skin aging. For instance, common signsof skin aging include, but are not limited to, appearance of fine linesor wrinkles, thin and transparent skin, loss of underlying fat (leadingto hollowed cheeks and eye sockets as well as noticeable loss offirmness on the hands and neck), bone loss (such that bones shrink awayfrom the skin due to bone loss, which causes sagging skin), dry skin(which might itch), inability to sweat sufficiently to cool the skin,unwanted facial hair, freckles, age spots, spider veins, rough andleathery skin, fine wrinkles that disappear when stretched, loose skin,or a blotchy complexion. According to the present disclosure, one ormore of the above signs of aging may be reduced, diminished, retarded oreven reversed by the compositions and methods of the present disclosure.

“Wound” means an injury to any tissue, including for example, acute,subacute, delayed or difficult to heal wounds, and chronic wounds.Examples of wounds may include both open and closed wounds. Woundsinclude, for example, amputations, burns, incisions, excisions, lesions,lacerations, abrasions, puncture or penetrating wounds, surgical wounds,contusions, hematomas, crushing injuries, ulcers (such as for examplepressure, diabetic or venous), wounds caused by periodontitis(inflammation of the periodontium).

Features and advantages of the subject matter hereof will become moreapparent in light of the following detailed description of selectedembodiments, as illustrated in the accompanying figures. As will berealized, the subject matter disclosed and claimed is capable ofmodifications in various respects, all without departing from the scopeof the claims. Accordingly, the drawings and the description are to beregarded as illustrative in nature, and not as restrictive and the fullscope of the subject matter is set forth in the claims.

(3) Biophotonic Materials

The present disclosure provides, in a broad sense, biophotonic materialswhich are cohesive and methods of using the biophotonic materials.Biophotonic materials can be, in a broad sense, activated by light(e.g., photons) of specific wavelength. A biophotonic material accordingto various embodiments of the present disclosure contains a cohesivematrix and at least one chromophore in or on the cohesive matrix whichis activated by light and accelerates the dispersion of light energy,which leads to light carrying on a therapeutic effect on its own, and/orto the photochemical activation of other agents contained in thecomposition (e.g., acceleration in the breakdown process of peroxide (anoxidant) when such compound is present in the composition or in contactwith the composition, leading to the formation of oxygen radicals, suchas singlet oxygen).

When a chromophore absorbs a photon of a certain wavelength, it becomesexcited. This is an unstable condition and the molecule tries to returnto the ground state, giving away the excess energy. For somechromophores, it is favorable to emit the excess energy as light whenreturning to the ground state. This process is called fluorescence. Thepeak wavelength of the emitted fluorescence is shifted towards longerwavelengths compared to the absorption wavelengths due to loss of energyin the conversion process. This is called the Stokes' shift. In theproper environment (e.g., in a biophotonic material) much of this energyis transferred to the other components of the biophotonic material or tothe treatment site directly.

Without being bound to theory, it is thought that fluorescent lightemitted by photoactivated chromophores may have therapeutic propertiesdue to its femto-, pico-, or nano-second emission properties which maybe recognized by biological cells and tissues, leading to favourablebiomodulation. Furthermore, the emitted fluorescent light has a longerwavelength and hence a deeper penetration into the tissue than theactivating light. Irradiating tissue with such a broad range ofwavelength, including in some embodiments the activating light whichpasses through the composition, may have different and complementaryeffects on the cells and tissues. In other words, chromophores are usedin the biophotonic materials of the present disclosure for therapeuticeffect on tissues. This is a distinct application of these photoactiveagents and differs from the use of chromophores as simple stains or ascatalysts for photo-polymerization.

The biophotonic materials of the present disclosure may have topicaluses such as a mask or a wound dressing, or as an attachment to a lightsource, as a waveguide or as a light filter. The cohesive nature ofthese biophotonic materials may provide ease of removal from the site oftreatment and hence a faster and less messy treatment. In addition thebiophotonic materials can limit the contact between the chromopore andthe tissue. These materials may be described based on the componentsmaking up the composition. Additionally or alternatively, thecompositions of the present disclosure have functional and structuralproperties and these properties may also be used to define and describethe compositions. Individual components of the biophotonic materials ofthe present disclosure, including chromophores, thickening agents andother optional ingredients, are detailed below.

The present disclosure also provides a precursor composition to thematerial described herein, which will become cohesive on drying,heating, light exposure, application to tissue or mixing. The precursorcomposition comprises at least one chromophore in a carrier medium, orat least one chromophore and a cohesive matrix.

(a) Chromophores

Suitable chromophores can be fluorescent dyes (or stains) (also known as“fluorochromes” or “fluorophores”). Other dye groups or dyes (biologicaland histological dyes, food colorings, carotenoids, naturally occurringfluorescent and other dyes) can also be used. Suitable photoactivatorscan be those that are Generally Regarded As Safe (GRAS). Advantageously,photoactivators which are not well tolerated by the skin or othertissues can be included in the biophotonic material of the presentdisclosure, as in certain embodiments, the photoactivators areencapsulated within the cohesive matrix and may not contact the tissues.

In certain embodiments, the biophotonic material of the presentdisclosure comprises a first chromophore which undergoes partial orcomplete photobleaching upon application of light. In some embodiments,the first chromophore absorbs at a wavelength in the range of thevisible spectrum, such as at a wavelength of about 380-800 nm, 380-700,400-800, or 380-600 nm. In other embodiments, the first chromophoreabsorbs at a wavelength of about 200-800 nm, 200-700 nm, 200-600 nm or200-500 nm. In one embodiment, the first chromophore absorbs at awavelength of about 200-600 nm. In some embodiments, the firstchromophore absorbs light at a wavelength of about 200-300 nm, 250-350nm, 300-400 nm, 350-450 nm, 400-500 nm, 450-650 nm, 600-700 nm, 650-750nm or 700-800 nm.

It will be appreciated to those skilled in the art that opticalproperties of a particular chromophore may vary depending on thechromophore's surrounding medium. Therefore, as used herein, aparticular chromophore's absorption and/or emission wavelength (orspectrum) corresponds to the wavelengths (or spectrum) measured in abiophotonic material of the present disclosure.

The biophotonic material disclosed herein may include at least oneadditional chromophore. Combining chromophores may increasephoto-absorption by the combined dye molecules and enhance absorptionand photo-biomodulation selectivity. This creates multiple possibilitiesof generating new photosensitive, and/or selective chromophoresmixtures. Thus, in certain embodiments, biophotonic materials of thedisclosure include more than one chromophore. When such multichromophorematerials are illuminated with light, energy transfer can occur betweenthe chromophores. This process, known as resonance energy transfer, is awidely prevalent photophysical process through which an excited ‘donor’chromophore (also referred to herein as first chromophore) transfers itsexcitation energy to an ‘acceptor’ chromophore (also referred to hereinas second chromophore). The efficiency and directedness of resonanceenergy transfer depends on the spectral features of donor and acceptorchromophores. In particular, the flow of energy between chromophores isdependent on a spectral overlap reflecting the relative positioning andshapes of the absorption and emission spectra. More specifically, forenergy transfer to occur, the emission spectrum of the donor chromophoremust overlap with the absorption spectrum of the acceptor chromophore(FIG. 1).

Energy transfer manifests itself through decrease or quenching of thedonor emission and a reduction of excited state lifetime accompaniedalso by an increase in acceptor emission intensity. FIG. 2 is aJablonski diagram that illustrates the coupled transitions involvedbetween a donor emission and acceptor absorbance.

To enhance the energy transfer efficiency, the donor chromophore shouldhave good abilities to absorb photons and emit photons. Furthermore, themore overlap there is between the donor chromophore's emission spectraand the acceptor chromophore's absorption spectra, the better a donorchromophore can transfer energy to the acceptor chromophore.

In certain embodiments, the biophotonic material of the presentdisclosure further comprises a second chromophore. In some embodiments,the first chromophore has an emission spectrum that overlaps at leastabout 80%, 50%, 40%, 30%, 20% or 10% with an absorption spectrum of thesecond chromophore. In one embodiment, the first chromophore has anemission spectrum that overlaps at least about 20% with an absorptionspectrum of the second chromophore. In some embodiments, the firstchromophore has an emission spectrum that overlaps at least 1-10%,5-15%, 10-20%, 15-25%, 20-30%, 25-35%, 30-40%, 35-45%, 50-60%, 55-65% or60-70% with an absorption spectrum of the second chromophore.

% spectral overlap, as used herein, means the % overlap of a donorchromophore's emission wavelength range with an acceptor chromophore'sabsorption wavelength rage, measured at spectral full width quartermaximum (FWQM). For example, FIG. 1 shows the normalized absorption andemission spectra of donor and acceptor chromophores. The spectral FWQMof the acceptor chromophore's absorption spectrum is from about 60 nm(515 nm to about 575 nm). The overlap of the donor chromophore'sspectrum with the absorption spectrum of the acceptor chromophore isabout 40 nm (from 515 nm to about 555 nm). Thus, the % overlap can becalculated as 40 nm/60 nm×100=66.6%.

In some embodiments, the second chromophore absorbs at a wavelength inthe range of the visible spectrum. In certain embodiments, the secondchromophore has an absorption wavelength that is relatively longer thanthat of the first chromophore within the range of about 50-250, 25-150or 10-100 nm.

The first chromophore can be present in an amount of about 0.001-40% perweight of the biophotonic material. When present, the second chromophorecan be present in an amount of about 0.001-40% per weight of thebiophotonic material. In certain embodiments, the first chromophore ispresent in an amount of about 0.001-3%, 0.001-0.01%, 0.005-0.1%,0.1-0.5%, 0.5-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%,15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%,32.5-37.5%, or 35-40% per weight of the biophotonic material. In certainembodiments, the second chromophore is present in an amount of about0.001-3%, 0.001-0.01%, 0.005-0.1%, 0.1-0.5%, 0.5-2%, 1-5%, 2.5-7.5%,5-10%, 7.5-12.5%, 10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%,22.5-27.5%, 25-30%, 27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40% per weightof the biophotonic material. In certain embodiments, the total weightper weight of chromophore or combination of chromophores may be in theamount of about 0.005-1%, 0.05-2%, 1-5%, 2.5-7.5%, 5-10%, 7.5-12.5%,10-15%, 12.5-17.5%, 15-20%, 17.5-22.5%, 20-25%, 22.5-27.5%, 25-30%,27.5-32.5%, 30-35%, 32.5-37.5%, or 35-40.001% per weight of thebiophotonic material.

The concentration of the chromophore to be used can be selected based onthe desired intensity and duration of the biophotonic activity from thebiophotonic material, and on the desired medical or cosmetic effect. Forexample, some dyes such as xanthene dyes reach a ‘saturationconcentration’ after which further increases in concentration do notprovide substantially higher emitted fluorescence. Further increasingthe chromophore concentration above the saturation concentration canreduce the amount of activating light passing through the matrix.Therefore, if more fluorescence is required for a certain applicationthan activating light, a high ‘saturation’ concentration of chromophorecan be used. However, if a balance is required between the emittedfluorescence and the activating light, a concentration close to or lowerthan the saturation concentration can be chosen.

Suitable chromophores that may be used in the biophotonic materials ofthe present disclosure include, but are not limited to the following:

Chlorophyll Dyes

Exemplary chlorophyll dyes include but are not limited to chlorophyll a;chlorophyll b; chlorophyllin, oil soluble chlorophyll;bacteriochlorophyll a; bacteriochlorophyll b; bacteriochlorophyll c;bacteriochlorophyll d; protochlorophyll; protochlorophyll a; amphiphilicchlorophyll derivative 1; and amphiphilic chlorophyll derivative 2.

Xanthene Derivatives

Exemplary xanthene dyes include but are not limited to Eosin B(4′,5′-dibromo,2′,7′-dinitr-o-fluorescein, dianion); eosin Y; eosin Y(2′,4′,5′,7′-tetrabromo-fluoresc-ein, dianion); eosin(2′,4′,5′,7′-tetrabromo-fluorescein, dianion); eosin(2′,4′,5′,7′-tetrabromo-fluorescein, dianion) methyl ester; eosin(2′,4′,5′,7′-tetrabromo-fluorescein, monoanion) p-isopropylbenzyl ester;eosin derivative (2′,7′-dibromo-fluorescein, dianion); eosin derivative(4′,5′-dibromo-fluorescein, dianion; eosin derivative(2′,7′-dichloro-fluorescein, danion); eosin derivative(4′,5′-dichloro-fluorescein, dianion); eosin derivative(2′,7′-diiodo-fluorescein, dianion); eosin derivative(4′,5′-diiodo-fluorescein, dianion); eosin derivative(tribromo-fluorescein, dianion); eosin derivative(2′,4′,5′,7′-tetrachlor-o-fluorescein, dianion); eosin; eosindicetylpyridinium chloride ion pair; erythrosin B(2′,4′,5′,7′-tetraiodo-fluorescein, dianion); erythrosin; erythrosindianion; erythiosin B; fluorescein; fluorescein dianion; phloxin B(2′,4′,5′,7′-tetrabromo-3,4,5,6-tetrachloro-fluorescein, dianion);phloxin B (tetrachloro-tetrabromo-fluorescein); phloxine B; rose bengal(3,4,5,6-tetrachloro-2′,4′,5′,7′-tetraiodofluorescein, dianion); pyroninG, pyronin J, pyronin Y; Rhodamine dyes such as rhodamines include4,5-dibromo-rhodamine methyl ester; 4,5-dibromo-rhodamine n-butyl ester;rhodamine 101 methyl ester; rhodamine 123; rhodamine 6G; rhodamine 6Ghexyl ester; tetrabromo-rhodamine 123; and tetramethyl-rhodamine ethylester.

Methylene Blue Dyes

Exemplary methylene blue derivatives include but are not limited to1-methyl methylene blue; 1,9-dimethyl methylene blue; methylene blue;methylene blue (16 μM); methylene blue (14 μM); methylene violet;bromomethylene violet; 4-iodoniethylene violet;1,9-dimethyl-3-dimethyl-amino-7-diethyl-a-mino-phenothiazine; and1,9-dimethyl-3-diethylamino-7-dibutyl-amino-phenot-hiazine.

Azo Dyes

Exemplary azo (or diazo-) dyes include but are not limited to methylviolet, neutral red, para red (pigment red 1), amaranth (Azorubine S),Carmoisine (azorubine, food red 3, acid red 14), allura red AC (FD&C40), tartrazine (FD&C Yellow 5), orange G (acid orange 10), Ponceau 4R(food red 7), methyl red (acid red 2), and murexide-ammonium purpurate.

In some aspects of the disclosure, the one or more chromophores of thebiophotonic materials disclosed herein can be independently selectedfrom any of Acid black 1, Acid blue 22, Acid blue 93, Acid fuchsin, Acidgreen, Acid green 1, Acid green 5, Acid magenta, Acid orange 10, Acidred 26, Acid red 29, Acid red 44, Acid red 51, Acid red 66, Acid red 87,Acid red 91, Acid red 92, Acid red 94, Acid red 101, Acid red 103, Acidroseine, Acid rubin, Acid violet 19, Acid yellow 1, Acid yellow 9, Acidyellow 23, Acid yellow 24, Acid yellow 36, Acid yellow 73, Acid yellowS, Acridine orange, Acriflavine, Alcian blue, Alcian yellow, Alcoholsoluble eosin, Alizarin, Alizarin blue 2RC, Alizarin carmine, Alizarincyanin BBS, Alizarol cyanin R, Alizarin red 5, Alizarin purpurin,Aluminon, Amido black 10B, Amidoschwarz, Aniline blue WS, Anthraceneblue SWR, Auramine O, Azocannine B, Azocarmine G, Azoic diazo 5, Azoicdiazo 48, Azure A, Azure B, Azure C, Basic blue 8, Basic blue 9, Basicblue 12, Basic blue 15, Basic blue 17, Basic blue 20, Basic blue 26,Basic brown 1, Basic fuchsin, Basic green 4, Basic orange 14, Basic red2, Basic red 5, Basic red 9, Basic violet 2, Basic violet 3, Basicviolet 4, Basic violet 10, Basic violet 14, Basic yellow 1, Basic yellow2, Biebrich scarlet, Bismarck brown Y, Brilliant crystal scarlet 6R,Calcium red, Carmine, Carminic acid, Celestine blue B, China blue,Cochineal, Coelestine blue, Chrome violet CG, Chromotrope 2R, Chromoxanecyanin R, Congo corinth, Congo red, Cotton blue, Cotton red, Croceinescarlet, Crocin, Crystal ponceau 6R, Crystal violet, Dahlia, Diamondgreen B, Direct blue 14, Direct blue 58, Direct red, Direct red 10,Direct red 28, Direct red 80, Direct yellow 7, Eosin B, Eosin Bluish,Eosin, Eosin Y, Eosin yellowish, Eosinol, Erie garnet B, Eriochromecyanin R, Erythrosin B, Ethyl eosin, Ethyl green, Ethyl violet, Evansblue, Fast blue B, Fast green FCF, Fast red B, Fast yellow, Fluorescein,Food green 3, Gallein, Gallamine blue, Gallocyanin, Gentian violet,Haematein, Haematine, Haematoxylin, Helio fast rubin BBL, Helvetia blue,Hematein, Hematine, Hematoxylin, Hoffman's violet, Imperial red,Indocyanin Green, Ingrain blue, Ingrain blue 1, Ingrain yellow 1, INT,Kermes, Kermesic acid, Kernechtrot, Lac, Laccaic acid, Lauth's violet,Light green, Lissamine green SF, Luxoi fast blue, Magenta 0, Magenta I,Magenta II, Magenta III, Malachite green, Manchester brown, Martiusyellow, Merbromin, Mercurochrome, Metanil yellow, Methylene azure A,Methylene azure B, Methylene azure C, Methylene blue, Methyl blue,Methyl green, Methyl violet, Methyl violet 2B, Methyl violet 10B,Mordant blue 3, Mordant blue 10, Mordant blue 14, Mordant blue 23,Mordant blue 32, Mordant blue 45, Mordant red 3, Mordant red 11, Mordantviolet 25, Mordant violet 39 Naphthol blue black, Naphthol green B,Naphthol yellow S, Natural black 1, Natural green 3(chlorophyllin),Natural red, Natural red 3, Natural red 4, Natural red 8, Natural red16, Natural red 25, Natural red 28, Natural yellow 6, NBT, Neutral red,New fuchsin, Niagara blue 313, Night blue, Nile blue, Nile blue A, Nileblue oxazone, Nile blue sulphate, Nile red, Nitro BT, Nitro bluetetrazolium, Nuclear fast red, Oil red O, Orange G, Orcein,Pararosanilin, Phloxine B, Picric acid, Ponceau 2R, Ponceau 6R, PonceauB, Ponceau de Xylidine, Ponceau S, Primula, Purpurin, Pyronin B,phycobilins, Phycocyanins, Phycoerythrins. Phycoerythrincyanin (PEC),Phthalocyanines, Pyronin G, Pyronin Y, Quinine, Rhodamine B, Rosanilin,Rose bengal, Saffron, Safranin O, Scarlet R, Scarlet red, Scharlach R,Shellac, Sirius red F3B, Solochrome cyanin R, Soluble blue, Solventblack 3, Solvent blue 38, Solvent red 23, Solvent red 24, Solvent red27, Solvent red 45, Solvent yellow 94, Spirit soluble eosin, Sudan III,Sudan IV, Sudan black B, Sulfur yellow S, Swiss blue, Tartrazine,Thioflavine S, Thioflavine T, Thionin, Toluidine blue, Toluyline red,Tropaeolin G, Trypaflavine, Trypan blue, Uranin, Victoria blue 4R,Victoria blue B, Victoria green B, Vitamin B, Water blue I, Watersoluble eosin, Xylidine ponceau, or Yellowish eosin.

In certain embodiments, the biophotonic material of the presentdisclosure includes any of the chromophores listed above, or acombination thereof, so as to provide a synergistic biophotonic effectat the application site.

Without being bound to any particular theory, a synergistic effect ofthe chromophore combinations means that the biophotonic effect isgreater than the sum of their individual effects. Advantageously, thismay translate to increased reactivity of the biophotonic material,faster or improved treatment time. Also, the treatment conditions neednot be altered to achieve the same or better reatment results, such astime of exposure to light, power of light source used, and wavelength oflight used. In other words, use of synergistic combinations ofchromophores may allow the same or better treatment withoutnecessitating a longer time of exposure to a light source, a higherpower light source or a light source with different wavelengths.

In some embodiments, the material includes Eosin Y as a firstchromophore and any one or more of Rose Bengal, Fluorescein, Erythrosin,Phloxine B, chlorophyllin as a second chromophore. It is believed thatthese combinations have a synergistic effect as they can transfer energyto one another the activated due in part to overlaps or close proximityof their absorption and emission spectra. This transferred energy isthen emitted as fluoresence or leads to production of reactive oxygenspecies. This absorbed and re-emitted light is thought to be transmittedthroughout the composition, and also to be transmitted into the site oftreatment.

In further embodiments, the material includes the following synergisticcombinations: Eosin Y and Fluorescein; Fluorescein and Rose Bengal;Erythrosine in combination with Eosin Y, Rose Bengal or Fluorescein;Phloxine B in combination with one or more of Eosin Y, Rose Bengal,Fluorescein and Erythrosine. Other synergistic chromophore combinationsare also possible.

By means of synergistic effects of the chromophore combinations in thematerial, chromophores which cannot normally be activated by anactivating light (such as a blue light from an LED), can be activatedthrough energy transfer from chromophores which are activated by theactivating light. In this way, the different properties ofphotoactivated chromophores can be harnessed and tailored according tothe cosmetic or the medical therapy required.

For example, Rose Bengal can generate a high yield of singlet oxygenwhen activated in the presence of molecular oxygen, however it has a lowquantum yield in terms of emitted fluorescent light. Rose Bengal has apeak absorption around 540 nm and so can be activated by green light.Eosin Y has a high quantum yield and can be activated by blue light. Bycombining Rose Bengal with Eosin Y, one obtains a composition which canemit therapeutic fluorescent light and generate singlet oxygen whenactivated by blue light. In this case, the blue light photoactivatesEosin Y which transfers some of its energy to Rose Bengal as well asemitting some energy as fluorescence.

In some embodiments, the chromophore or chromophores are selected suchthat their emitted fluorescent light, on photoactivation, is within oneor more of the green, yellow, orange, red and infrared portions of theelectromagnetic spectrum, for example having a peak wavelength withinthe range of about 490 nm to about 800 nm. In certain embodiments, theemitted fluorescent light has a power density of between 0.005 to about10 mW/cm², about 0.5 to about 5 mW/cm².

(b) Cohesive Matrix

The biophotonic material of the present disclosure comprise a cohesivematrix made from one or more thickening agents, or a carrier medium. Inother words, the biophotonic material of the present disclosure compriseone or more thickening agents, or a carrier medium which can form acohesive matrix. These agents are present in an amount and ratiosufficient to provide a desired viscosity, flexibility, rigidity,tensile strength, tear strength, elasticity, and adhesiveness. Thedesired properties may be one of achieving a peelable film, or a rigidor flexible matrix. The thickening agents are selected so that thechromophore can remain photoactive in the cohesive matrix. Thethickening agents are also selected according to the opticaltransparency of the cohesive matrix which they will form. The cohesivematrix should be able to transmit sufficient light to activate the atleast one chromophore and, in embodiments where fluorescence is emittedby the activated chromophore, the cohesive matrix should also be able totransmit the emitted fluorescent light to tissues. It will be recognizedby persons skilled in the art that the thickening agent is anappropriate medium for the chromophore selected. For example, sonicxanthene dyes do not fluoresce in non-hydrated media, so hydratedpolymers or polar solvents may be used. The thickening agents shouldalso be selected according to the intended use. For example, if thebiophotonic material is to be applied onto tissue, the thickening agentis preferably a biocompatible material, or the cohesive matrix has anoutside layer of a biocompatible material which will interface thetissue.

Thickening Agents

In some embodiments, the content of a thickening agent used to make thecohesive matrix is from about 0.001% to about 40% (w/w %) of the totalweight. In certain embodiments, the total content of the thickeningagent is about 0.001-0.01%, about 0.005-0.05%, about 0.01-0.1, about0.05-0.5% about 0.1-1%, about 0.5-5%, about 1-5%, about 2.5-7.5%, about5-10%, about 7.5-12.5%, about 10-15%, about 12.5-17.5%, or about 15-20%,or about 15-25%, or about 20-30%, or about 25-35%, or about 30-40%. Itwill be recognized by one of skill in the art that the viscosity,flexibility, rigidity, tensile strength, tear strength, elasticity, andadhesiveness can be adjusted by varying the content of the thickeningmaterial. Methods of determining viscosity, flexibility, rigidity,tensile strength, tear strength, elasticity, and adhesiveness are knownin the art.

Thickening agents that can be used to prepare the biophotonic materialsof the present disclosure include polymers, copolymers, and monomers of:vinylpyrrolidones, methacrylamides, acrylamides N-vinylimidazoles,carboxy vinyls, vinyl esters, vinyl ethers, silicones,polyethyleneoxides, polyethyleneglycols, vinylalcohols, sodiumacrylates, acrylates, maleic acids, NN-dimethylacrylamides, diacet neacrylamides, acrylamides, acryloyl morpholine, pluronic, collagens,polyacrylamides, polyacrylates, polyvinyl alcohols, polyvinylenes,polyvinyl silicates, polyacrylates substituted with a sugar (e.g.,sucrose, glucose, glucosamines, galactose, trehalose, mannose, orlactose), acylamidopropane sulfonic acids, tetramethoxyorthosilicates,methyltrimethoxyorthosilicates, tetraalkoxyorthosilicates,trialkoxyorthosilicates, glycols, propylene glycol, glycerine,polysaccharides, alginates, dextrans, cyclodextrin, celluloses, modifiedcelluloses, oxidized celluloses, chitosans, chitins, guars,carrageenans, hyaluronic acids, inulin, starches, modified starches,agarose, methylcelluloses, plant gums, hylaronans, hydrogels, gelatins,glycosaminoglycans, carboxymethyl celluloses, hydroxyethyl celluloses,hydroxy propyl methyl celluloses, pectins, low-methoxy pectins,cross-linked dextrans, starch-acrylonitrile graft copolymers, starchsodium polyacrylate, hydroxyethyl methacrylates, hydroxyl ethylacrylates, polyvinylene, polyethylvinylethers, polytnethylmethacrylates, polystyrenes, polyurethanes, polyalkanoates, polylacticacids, polylactates, poly(3-hydroxybutyrate), sulfonated hydrogels, AMPS(2-acrylamide-2-methyl-1-propanesulfonic acid), SEM(sulfoethylmethacrylate), SPM (sulfopropyl methacrylate), SPA(sulfopropyl acrylate),N,N-dimethyl-N-tnethacryloxyethyl-N-(3-sulfoproyl)ammonium betaine,methacryllic acid amidopropyl-dimethyl ammonium sulfobetaine, SPI{itaconic acid-bis(1-propyl sulfonizacid-3) ester di-potassium salt},itaconic acids, AMBC (3-acrylamido-3-methylbutanoic acid),beta-carboxyethyl acrylate (acrylic acid dimers), and maleicanhydride-methylvinyl ether polymers, derivatives thereof, saltsthereof, acids thereof, combinations thereof, and the like,

Thickening agents also include poly (ethylene oxide) polymers (such asPOLYOX from Dow Chemical), linear PVP and cross-linked PVP, PEG/PPGcopolymers (such as BASF Pluracare L1220), ethylene oxide (EO)-propyleneoxide (PO) block copolymers (such as polymers sold under the trade markPluronic available from BASF Corporation), ester gum, shellac, pressuresensitive silicone adhesives (such as BioPSA from Dow-Corning), ormixtures thereof. In some embodiments, a copolymer comprises (PVM/MA).In an embodiment, a copolymer comprises poly (methylvinylether/maleicanhydride). In some embodiments, a copolymer comprises poly(methylvinylether/maleic acid). In some embodiments, a copolymercomprises poly (methylvinylether/maleic acid) half esters. In someembodiments, a copolymer comprises poly (methylvinylether/maleic acid)mixed salts.

Thickening agents can also include carbomers which are a synthetic highmolecular weight polymer of acrylic acid that is crosslinked with eitherallylsucrose or allylethers of pentaerythritol having a molecular weightof about 3×10⁶. The gelation mechanism depends on neutralization of thecarboxylic acid moiety to form a soluble salt. The polymer ishydrophilic and produces sparkling clear gels when neutralized.Carbomers are available as fine white powders which disperse in water toform acidic colloidal suspensions (a 1% dispersion has approx. pH 3) oflow viscosity. Neutralization of these suspensions using a base, forexample sodium, potassium or ammonium hydroxides, low molecular weightamines and alkanolamines, results in the formation of clear translucentgels.

In one embodiment of the disclosure, the carbomer is Carbopol. Suchpolymers are commercially available from B.F. Goodrich or Lubrizol underthe designation Carbopol® 71G N.F, 420, 430, 475, 488, 493, 910, 934,934P, 940, 971PNF, 974P NF, 980 NF, 981 NF and the like. Carbopols areversatile controlled-release polymers, as described by Brock(Pharmacotherapy, 14:430-7 (1994)) and Durrani (Pharmaceutical Res.(Supp.) 8:S-135 (1991)), and belong to a family of carbomers which aresynthetic, high molecular weight, non-linear polymers of acrylic acid,cross-linked with polyalkenyl polyether. In some embodiments, thecarbomer is Carbopol® 974P NF, 980 NF, 5984 EP, ETD 2020NF, Ultrez 10NF, 934 NF, 934P NF or 940 NE. In certain embodiments, the carbomer isCarbopol® 980 NF, ETD 2020 NF, Ultrez 10 NF, Ultrez 21 or 1382 Polymer,1342 NE, 940 NF.

The biophotonic material of the present disclosure may be water soluble.Alternatively, the biophotonic material of the present disclosure mayoptionally include a water-insoluble substrate. By “water insoluble”, itis meant that the substrate does not dissolve in or readily break apartupon immersion in water. In some embodiments, the water-insolublesubstrate is the implement or vehicle for delivering the treatmentcomposition to the skin or target tissue. A wide variety of materialscan be used as the water-insoluble substrate. The following non-limitingcharacteristics are desirable: (i) sufficient wet strength for use, (ii)sufficient softness, (iii) sufficient thickness, (iv) appropriate size,(v) air permeability, and (vi) hydrophilicity.

Non-limiting examples of suitable water-insoluble substrates which meetthe above criteria include nonwoven substrates, woven substrates,hydroentangled substrates, air entangled substrates, natural sponges,synthetic sponges, polymeric netted meshes, and the like. Preferredembodiments employ nonwoven substrates since they are economical andreadily available in a variety of materials. By “nonwoven”, it is meantthat the layer is comprised of fibers which are not woven into a fabricbut rather are formed into a sheet, mat, or pad layer.

Antimicrobials

Antimicrobials kill microbes or inhibit their growth or accumulation,and are optionally included in the biophotonic materials of the presentdisclosure, Exemplary antimicrobials (or antimicrobial agent) arerecited in U.S. Patent Application Publications 20040009227 and20110081530. Suitable antimicrobials for use in the methods andcompositions of the present disclosure include, but not limited to,hydrogen peroxide, urea hydrogen peroxide, benzoyl peroxide, phenolicand chlorinated phenolic and chlorinated phenolic compounds, resorcinoland its derivatives, bisphenolic compounds, benzoic esters (parabens),halogenated cathonilides, polymeric antimicrobial agents, thazolines,trichloromethylthioimides, natural antimicrobial agents (also referredto as “natural essential oils”), metal salts, and broad-spectrumantibiotics.

Hydrogen peroxide (H₂O₂) is a powerful oxidizing agent, and breaks downinto water and oxygen and does not form any persistent, toxic residualcompound. A suitable range of concentration over which hydrogen peroxidecan be used in the biophotonic material is from about 0.1% to about 3%,about 0.1 to 1.5%, about 1%.

Urea hydrogen peroxide (also known as urea peroxide, carbamide peroxideor percarbamide) is soluble in water and contains approximately 35%hydrogen peroxide. A suitable range of concentration over which ureaperoxide can be used in the biophotonic material of the presentdisclosure is from about 0.3% to about 5%. Urea peroxide breaks down tourea and hydrogen peroxide in a slow-release fashion that can beaccelerated with heat or photochemical reactions.

Benzoyl peroxide consists of two benzoyl groups (benzoic acid with the Hof the carboxylic acid removed) joined by a peroxide group. It is foundin treatments for acne, in concentrations varying from 2.5% to 10%. Thereleased peroxide groups are effective at killing bacteria. Benzoylperoxide also promotes skin turnover and clearing of pores, whichfurther contributes to decreasing bacterial counts and reduce acne.Benzoyl peroxide breaks down to benzoic acid and oxygen upon contactwith skin, neither of which is toxic. A suitable range of concentrationover which benzoyl peroxide can be used in the matrix biophotonic isfrom about 2.5% to about 5%.

According to certain embodiments, the biophotonic material of thepresent disclosure may optionally comprise one or more additionalcomponents, such as oxygen-rich compounds as a source of oxygenradicals. Peroxide compounds are oxidants that contain the peroxy group(R—O—O—R), which is a chainlike structure containing two oxygen atoms,each of which is bonded to the other and a radical or some element. Whena biophotonic material of the present disclosure comprising an oxidantis illuminated with light, the chromophores are excited to a higherenergy state. When the chromophores' electrons return to a lower energystate, they emit photons with a lower energy level, thus causing theemission of light of a longer wavelength (Stokes' shift). In the properenvironment, some of this energy is transferred to oxygen or thereactive hydrogen peroxide and causes the formation of oxygen radicals,such as singlet oxygen. The singlet oxygen and other reactive oxygenspecies generated by the activation of the biophotonic material arethought to operate in a hormetic fashion. That is, a health beneficialeffect that is brought about by the low exposure to a normally toxicstimuli (e.g. reactive oxygen), by stimulating and modulating stressresponse pathways in cells of the targeted tissues. Endogenous responseto exogenous generated free radicals (reactive oxygen species) ismodulated in increased defense capacity against the exogenous freeradicals and induces acceleration of healing and regenerative processes.Furthermore, activation of the oxidant will also produce anantibacterial effect. The extreme sensitivity of bacteria to exposure tofree radicals makes the biophotonic material of the present disclosure ade facto bactericidal composition.

Specific phenolic and chlorinated phenolic antimicrobial agents that canbe used in the disclosure include, but are not limited to: phenol;2-methyl phenol; 3-methyl phenol; 4-methyl phenol; 4-ethyl phenol;2,4-dimethyl phenol; 2,5-dimethyl phenol; 3,4-dimethyl phenol;2,6-dimethyl phenol; 4-n-propyl phenol; 4-n-butyl phenol; 4-n-amylphenol; 4-tert-amyl phenol; 4-n-hexyl phenol; 4-n-heptyl phenol; mono-and poly-alkyl and aromatic halophenols; p-chlorophenyl; methylp-chlorophenol; ethyl p-chlorophenol; n-propyl p-chlorophenol; n-butylp-chlorophenol; n-amyl p-chlorophenol; sec-amyl p-chlorophenol; n-hexylp-chlorophenol; cyclohexyl p-chlorophenol; n-heptyl p-chlorophenol;n-octyl; p-chlorophenol; o-chlorophenol; methyl o-chlorophenol; ethylo-chlorophenol; n-propyl o-chlorophenol; n-butyl o-chlorophenol; n-amylo-chlorophenol; tert-amyl o-chlorophenol; n-hexyl o-chlorophenol;n-heptyl o-chlorophenol; o-benzyl p-chlorophenol; o-benxyl-m-methylp-chlorophenol; o-benzyl-m,m-dimethyl p-chlorophenol; o-phenylethylp-chlorophenol; o-phenylethyl-m-methyl p-chlorophenol; 3-methylp-chlorophenol 3,5-dimethyl p-chlorophenol, 6-ethyl-3-methylp-chlorophenol, 6-n-propyl-3-methyl p-chlorophenol;6-iso-propyl-3-methyl p-chlorophenol; 2-ethyl-3,5-dimethylp-chlorophenol; 6-sec-butyl-3-methyl p-chlorophenol;2-iso-propyl-3,5-dimethyl p-chlorophenol; 6-diethylmethyl-3-methylp-chlorophenol; 6-iso-propyl-2-ethyl-3-methyl p-chlorophenol;2-sec-amyl-3,5-dimethyl p-chlorophenol; 2-diethylmethyl-3,5-dimethylp-chlorophenol; 6-sec-octyl-3-methyl p-chlorophenol; p-chloro-m-cresolp-bromophenol; methyl p-bromophenol; ethyl p-bromophenol; n-propylp-bromophenol; n-butyl p-bromophenol; n-amyl p-bromophenol; sec-amylp-bromophenol; n-hexyl p-bromophenol; cyclohexyl p-bromophenol;o-bromophenol; tert-amyl o-bromophenol; n-hexyl o-bromophenol;n-propyl-m,m-dimethyl o-bromophenol; 2-phenyl phenol; 4-chloro-2-methylphenol; 4-chloro-3-methyl phenol; 4-chloro-3,5-dimethyl phenol;2,4-dichloro-3,5-dimethylphenol; 3,4,5_(;)6-tetabromo-2-methylphenol-;5-methyl-2-pentylphenol; 4-isopropyl-3-methylphenol;para-chloro-metaxylenol (PCMX); chlorothymol; phenoxyethanol;phenoxyisopropanol; and 5-chloro-2-hydroxydiphenylmethane.

Resorcinol and its derivatives can also be used as antimicrobial agents.Specific resorcinol derivatives include but are not limited to: methylresorcinol; ethyl resorcinol; n-propyl resorcinol; n-butyl resorcinol;n-atnyl resorcinol; n-hexyl resorcinol; n-heptyl resorcinol; n-octylresorcinol; n-nonyl resorcinol; phenyl resorcinol; benzyl resorcinol;phenylethyl resorcinol; phenylpropyl resorcinol; p-chlorobenzylresorcinol; 5-chloro-2,4-dihydroxydiphenyl methane;4′-chloro-2,4-dihydroxydiphenyl methane; 5-bromo-2,4-dihydroxydiphenylmethane; and 4′-bromo-2,4-dihydroxydiphenyl methane. Specificbisphenolic antimicrobial agents that can be used in the disclosureinclude, but are not limited to: 2,2′-methylene bis-(4-chlorophenol);,4,4′trichloro-2′-hydroxy-diphenyl ether, which is sold by Ciba Geigy,Florham Park, N.J. under the tradename Triclosan®; 2,2′-methylenebis-(3,4,6-trichlorophenol); 2,2′-methylenebis-(4-chloro-6-bromophenol); bis-(2-hydroxy-3,5-dichlorop- hexyl)sulphide; and bis-(2-hydroxy-5-chlorobenzyl)sulphide. Specific benzoieesters (parabens) that can be used in the disclosure include, but arenot limited to: methylparaben; propylparaben; butylparaben;ethylparaben; isopropylparaben; isobutylparaben; benzylparaben; sodiummethylparaben; and sodium propylparaben.

Specific halogenated carbanilides that can be used in the disclosureinclude, but are not limited to: 3,4,4′-trichlorocarbanilides, such as3-(4-chlorophenyl)-1-(3,4-dichlorpheny)urea sold under the tradenameTriclocarban® by Ciba-Geigy, Florham Park, N.J.;3-trifluoromethyl-4,4′-dichlorocarbanilide; and3,3′,4-trichlorocarbanilide. Specific polymeric antimicrobial agentsthat can be used in the disclosure include, but are not limited to:polyhexamethylene biguanide hydrochloride; and poly(iminoimidocarbonyliminoimidocarbonyl iminohexamethylene hydrochloride), which is soldunder the tradename Vantocil® IB.

Specific thazolines that can be used in the disclosure include, but arenot limited to that sold under the tradename Micro-Check®; and2-n-octyl-4-isothiazolin-3-one, which is sold under the tradenameVinyzene® IT-3000 DIDP. Specific trichlorornethylthioimides that can beused in the disclosure include, but are not limited to:N-(trichloromethylthio)phthalimide, which is sold under the tradenameFungitrol®; and N-trichloromethylthio-4-cyclohexene-1,2-dicarboximide,which is sold under the tradename Vancide®.

Specific natural antimicrobial agents that can be used in the disclosureinclude, but are not limited to, oils of: anise; lemon; orange;rosemary; wintergreen; thyme; lavender; cloves; hops; tea tree;citronella; wheat; barley; lemongrass; cedar leaf; cedarwood; cinnamon;fleagrass; geranium; sandalwood; violet; cranberry; eucalyptus; vervain;peppermint; gum benzoin; basil; fennel; fir; balsam; menthol; ocmeaoriganuin; hydastis; carradensis; Berberidaceac daceae; Ratanhiae longa;and Curcuma longa. Also included in this class of natural antimicrobialagents are the key chemical components of the plant oils which have beenfound to provide antimicrobial benefit. These chemicals include, but arenot limited to: anethol; catechole; camphene; thymol; eugenol;eucalyptol; ferulic acid; farnesol; hinokitiol; tropolone; limonene;menthol; methyl salicylate; carvacol; terpineol; verbenone; berberine;ratanhiae extract; caryophellene oxide; citronellic acid; curcumin;nerolidol; and geraniol.

Specific metal salts that can be used in the disclosure include, but arenot limited to, salts of metals in groups 3a-5a, 3b-7b, and 8 of theperiodic table. Specific examples of metal salts include, but are notlimited to, salts of: aluminum; zirconium; zinc; silver; gold; copper;lanthanum; tin; mercury; bismuth; selenium; strontium; scandium;yttrium; cerium; praseodymiun; neodymium; promethum; samarium; europium;gadolinium; terbium; dysprosium; holmium; erbium; thalium; ytterbium;lutetium; and mixtures thereof. An example of the metal-ion basedantimicrobial agent is sold under the tradename HealthShield®, and ismanufactured by HealthShield Technology, Wakefield, Mass.

Specific broad-spectrum antimicrobial agents that can be used in thedisclosure include, but are not limited to, those that are recited inother categories of antimicrobial agents herein.

Additional antimicrobial agents that can be used in the methods of thedisclosure include, but are not limited to: pyrithiones, and inparticular pyrithione-including zinc complexes such as that sold underthe tradename Octopirox®; dimethyidimethylol hydantoin, which is soldunder the tradename Glydant®; methylchloroisothiazolinone/methylisothiazolinone, which is sold under thetradename Kathon CG®; sodium sulfite; sodium bisulfite; imidazolidinylurea, which is sold under the tradename Germall 115®; diazolidinyl urea,which is sold under the tradename Germall 11®;benzyl alcoholv2-bromo-2-nitropropane-1,3-diol, which is sold under the tradenameBronopol®; formalin or formaldehyde; iodopropenyl butylcarbamate, whichis sold under the tradename Polyphase P1000®; chloroacetamide;methanamine; methyldibromonitrile glutaronitrile(1,2-dibromo-2,4-dicyanobutane), which is sold under the tradenameTektamer®; glutaraldehyde; 5-bromo-5-nitro-1,3-dioxane, which is soldunder the tradename Bronidox®; phenethyl alcohol; o-phenylphenol/sodiumo-phenylphenol sodium hydroxymethylglycinate, which is sold under thetradename Suttocide A®; polymethoxy bicyclic oxazolidine; which is soldunder the tradename Nuosept C®; dimethoxane; thimersal; dichlorobenzylalcohol; captan; chlorphenenesin; dichlorophene; chlorbutanol; glyceryllaurate; halogenated diphenyl ethers;2,4,4′-trichloro-2′-hydroxy-diphenyl ether, which is sold under thetradename Triclosan® and is available from Ciba-Geigy, Florham Park,N.J.; and 2,2′-dihydroxy-5,5′-dibromo-diphenyl ether.

Additional antimicrobial agents that can be used in the methods of thedisclosure include those disclosed by U.S. Pat. Nos. 3,141,321;4,402,959; 4,430,381; 4,533,435; 4,625,026; 4,736,467; 4,855,139;5,069,907; 5,091,102; 5,639,464; 5,853,883; 5,854,147; 5,894,042; and5,919,554, and U.S. Pat, Appl. Publ. Nos. 20040009227 and 20110081530,

(4) Optical Properties of the Biophotonic Materials

In certain embodiments, biophotonic materials of the present disclosureare substantially transparent or translucent. The % transmittance of thebiophotonic material can be measured in the range of wavelengths from250 nm to 800 nm using, for example, a Perkin-Elmer Lambda 9500 seriesUV-visible spectrophotometer. In some embodiments, transmittance at 460nm is measured. As transmittance is dependent upon thickness, thethickness of each sample can be measured with calipers prior to loadingin the spectrophotometer. Transmittance values can be normalized to athickness of 100 um (or any thickness) according to

${{F_{T - {corr}}\left( {\lambda,t_{2}} \right)} = {\left\lbrack {{^{- \sigma_{t}}(\lambda)}t_{1}} \right\rbrack^{\frac{t_{2}}{t_{1}}} = \left\lbrack {F_{T - {corr}}\left( {\lambda,t_{1}} \right)} \right\rbrack^{\frac{t_{2}}{t_{1}}}}},$

where t₁=actual specimen thickness, t₂=thickness to which transmittancemeasurements can be normalized.

In certain embodiments, the biophotonic materials are substantiallyopaque. In these embodiments, the biophotonic materials may includelight transmitting structures such as fibres, particles, networks, whichare made of materials which can transmit light. The light transmittingstructures can be waveguides such as optical fibres.

In some embodiments, the biophotonic material has a transmittance thatis more than about 20%, 30%, 40%, 50%, 60%, 70%, or 75% at 460 nm. Insome embodiments, the transmittance exceeds 40% at 460 nm, 41% at 460nm, 42% at 460 nm, 43% at 460 nm, 44% at 460 nm, or 45% at 460 nm.

(5) Forms of the Biophotonie Materials

The biophotonic materials of the present disclosure may be in the formof a cohesive film or matrix containing at least one chromophore. Thecohesive film or matrix may be a cohesive gel, or a paste, a putty, asemi-solid, or a solid.

The biophotonic materials of the present disclosure may be deformable.They may be elastic or non-elastic. The biophotonic materials, forexample, may be in a peel-off form (peelable') to provide ease and speedof use. In certain embodiments, the tear strength and/or tensilestrength of the peel-off form is greater than its adhesion strength.This may help handleability of the material. It will be recognized byone of skill in the art that the properties of the peel-off biophotonicmaterial such as cohesiveness, flexibility, elasticity, tensilestrength, and tearing strength, can be determined and/or adjusted bymethods known in the art such as by selecting suitable thickening agentsand adapting their relative ratios.

The biophotonic material may be in a pre-formed shape. In certainembodiments, the pre-formed shape is in the form of, including, but notlimited to, a film, a face mask, a patch, or bandage. In certainembodiments, the pre-formed shapes can be customized for the individualuser by trimming to size. In certain embodiments, perforations areprovided around the perimeter of the pre-formed shape to facilitatetrimming.

A biophotonic material of the disclosure can be configured with a shapeand/or size for application to a desired portion of a subject's body.For example, the biophotonic material can be shaped and sized tocorrespond with a desired portion of the body to receive the biophotonictreatment. Such a desired portion of skin can be selected from, but notlimited to, the group consisting of a skin, head, forehead, scalp, nose,cheeks, lips, ears, face, neck, shoulder, arm pit, arm, elbow, hand,finger, abdomen, chest, stomach, back, buttocks, sacrum, genitals, legs,knee, feet, toes, nails, hair, any boney prominences, and combinationsthereof, and the like. Thus, the biophotonic material of the disclosurecan be shaped and sized to be applied to any portion of skin on asubject's body. For example, the biophotonic material can be sock, hat,glove or mitten shaped. In embodiments where the biophotonic material iselastic or rigid, it can be peeled-off without leaving any residue onthe tissue.

In certain embodiments, the biophotonic material is in the form of a anelastic and peelable face mask, which may be pre-formed. In otherembodiments, the biophotonic material is in the form of a non-elasticface mask, which may also be pre-formed. The mask can have openings forone or more of the eyes, nose and mouth. In a further embodiment, theopenings are protected with a covering, or the exposed skin such as onthe nose, lips or eyes are protected using for example cocoa butter. Incertain embodiments, the pre-formed face mask is provided in the form ofmultiple parts, e.g., an upper face part and a lower face part. Incertain embodiments, the uneven proximity of the face to a light sourceis compensated for, e.g., by adjusting the thickness of the mask, or byadjusting the amount of chromophore in the different areas of the mask,or by blocking the skin in closest proximity to the light. In certainembodiments, the pre-formed shapes come in a one-size fits all form.

In certain aspects, the mask (or patch) is not pre-formed and is appliede.g., by spreading a composition making up the mask (or patch), on theskin or target tissue, or alternatively by spraying, smearing, dabbingor rolling the composition on target tissue. It can then be converted toa peel-off form after application, by means such as, but not limited to,drying, illumination with light, change in temperature or pH uponapplication to the skin or tissue. The mask (or patch) can then bepeeled off without leaving any flakes on the skin or tissue, preferablywithout wiping or washing.

In certain aspects, the biophotonic material may have shape memoryproperties. For example, the biophotonic material can include a shapememory material, such as a shape memory polymer whose original shape isreverted to on activation by light. The original shape can be a flat orconcave configuration which allows the film/matrix to be readily peeledoff the tissue. The shape memory material may be included as a layerattached to the biophotonic material, or integrated with the biophotonicmaterial.

In certain aspects, the biophotonic material forms part of a compositeand can include fibres, particulates, non-biophotonic layers orbiophotonic layers with the same or different compositions.

In certain embodiments, the biophotonic material may comprise aplurality of waveguides extending at least partially through thebiophotonic material or contained at least partially within thebiophotonic material. The waveguides can be attached to a light sourceto thereby illuminate the biophotonic material from within. Thebiophotonic material may further include the light source attached tothe waveguides. The waveguides can be optical fibres which can transmitlight, not only from their ends, but also from their body. For example,made of polycarbonate or polymethylmetacrylate or any other suitablematerial.

In a different embodiment, the biophotonic material comprises a layer ofa woven or non-woven fabric dressing or a mask. Waveguides or a lightsource may be included within the dressing or mask fabric. For example,the dressing or mask fabric can be in the form of an envelope whichreceives the biophotonic material, and which comprises at least onelight emitting surface.

In certain aspects, the biophotonic material is formed as a filter. Forexample, the biophotonic material can be made to have a shape and a sizewhich can be connected to, or spaced from, a light emitting surface of alamp. In one embodiment, the lamp can be a hand-held lamp such as atorch or a dentist's curing lamp. The lamp with the biophotonic filtercan then be used to treat tissue sites of patient in a contacting ornon-contacting manner. In this embodiment, the filter has a body havinga first end which is sized and shaped to be connectable to a lightemitting surface, and a second end shaped to treat tissues.

In certain aspects, the biophotonic material is formed as a waveguide.In certain embodiments, at least one chromophore is included in anelongate solid matrix having good light propagation properties andappropriate mechanical properties. The waveguide may be flexible. Thewaveguide can be shaped as an optical fibre. Such an optical fibre canbe connected to a light source, and the at least one chromophore in thecohesive matrix activated by the light source to deliver therapeuticfluorescent light to hard to reach places, such as internal cavities andperiodontal pockets. Polymethylmethacrylate is an example of anappropriate cohesive matrix for use as a biophotonic waveguide. Such awaveguide may additionally include a coating to prevent lightdissipation from along its length.

In other aspects, the biophotonic material comprising at least onechromophore and a cohesive matrix is in the form of particulates.Material processing techniques known in the art can be used to formparticulates of any shape or size. These particulates can be containedin semi-solid or liquid preparations. For example, such biophotonicparticulates can be used in skin preparations such as creams, emulsionsto provide therapeutic effect to the skin. In this case, a biocompatiblesolid matrix is used and can be used to encapsulate all types ofchromophores, even those not well tolerated by the skin.

The biophotonic materials of the present disclosure may have a thicknessof from about 0.1 mm to about 50 mm. It will be appreciated that thethickness of the biophotonic materials will vary based on the intendeduse. In some embodiments, the biophotonic material has a thickness offrom about 0.1-1 mm. In some embodiments, the biophotonic material has athickness of about 0.5-1.5 mm, about 1-2 mm, about 1.5-2.5 mm, about 2-3mm, about 2.5-3.5 mm, about 3-4 mm, about 3.5-4.5 mm, about 4-5 mm,about 4.5-5.5 mm, about 5-6 mm, about 5.5-6.5 mm, about 6-7 mm, about6.5-7.5 mm, about 7-8 mm, about 7.5-8.5 mm, about 8-9 min, about8.5-9.5, about 9-10 mm, about 10-11 mm, about 11-12 mm, about 12-13 mm,about 13-14 mm, about 14-15 mm, about 15-16 mm, about 16-17 mm, about17-18 mm, about 18-19 mm, about 19-20 mm, about 20-22mm, about 22-24mm,about 24-26mm, about 26-28mm, about 28-30mm, about 30-35mm, about35-40mm, about 40-45mm, about 45-50mm.

The tensile strength of the biophotonic materials will vary based on theintended use. The tensile strength can be determined by performing atensile test and recording the force and displacement. These are thenconverted to stress (using cross sectional area) and strain; the highestpoint of the stress-strain curve is the “ultimate tensile strength.” Insome embodiments, tensile strength can be characterized using a 500Ncapacity tabletop mechanical testing system (#5942R4910, Instron) with a5N maximum static load cell (#102608, Instron). Pneumatic side actiongrips can be used to secure the samples (#2712-019, Instron). In someembodiments, a constant extension rate (for example, of about 2 mm/min)until failure can be applied and the tensile strength is calculated fromthe stress vs. strain data plots.

In some embodiments, the biophotonic material has a tensile strength offrom about 50 kPa to about 600 kPa. In some embodiments, the tensilestrength is from about 75 kPa to about 500 kPa, from about 100 kPa toabout 200 kPa, 100-300 kPa, 400 kPa, from about 150 kPa to about 350kPa, or from about 200 kPa to about 300 kPa.

In some embodiments, the tensile strength is at least about 50 kPa, atleast about 75 kPa, at least about 100 kPa, at least about 150 kPa, atleast about 200 kPa, at least about 250 kPa, at least about 300 kPa, atleast about 350 kPa, at least about 400 kPa, at least about 450 kPa, atleast about 500 kPa, at least about 550 kPa or at least about 600 kPa.

The tear strength of the biophotonic material will vary depending on theintended use. The tear strength property of the biophotonic material canbe tested using a 500N capacity tabletop mechanical testing system(#5942R4910, Instron) with a 5N maximum static load cell (#102608Instron). Pneumatic side action grips can be used to secure the samples(#2712-019, Instron). Samples can be tested with a constant extensionrate (for example, of about 2 mm/min) until failure. In accordance withthe invention, tear strength is calculated as the force at failuredivided by the average thickness (N/mm).

In some embodiments, the biophotonic material has a tear strength offrom about 0.1 N/mm to about 1 N/mm. In some embodiments, the tearstrength is from about 0.20 N/mm to about 0.40 N/mm, from about 0.25N/mn to about 0.35 N/mm, from about 0.25 N/mm to about 0.45 N/mm, fromabout 0.35 N/mm to about 0.535 N/mm, from about 0.45 N/mm to about 0.65N/mm, from about 0.55 N/mm to about 0.75 N/mm, from about 0.65 N/mm toabout 0.85 N/mm, from about 0.75 N/mm to about 0.95 N/mm.

In some embodiments, the tear strength is at least about 0.10 N/mm, atleast about 0.15 N/mm, at least about 0.20 N/mm, at least about 0.25N/mm, at least about 0.30 N/mm, at least about 0.35 N/mm, at least about0.40 N/mm, at least about 0.45 N/mm, at least about 0.55 N/mm or atleast about 1 N/mm.

The adhesion strength of the biophotonic material will vary depending onthe intended use. Adhesion strength is determined in accordance withASTM D-3330-78, PSTC-101 and is a measure of the force required toremove a biophotonic material from a test panel at a specific angle andrate of removal. A predetermined size of a biophotonic material of isapplied to a horizontal surface of a clean glass test plate. A hardrubber roller is used to firmly apply the piece and remove alldiscontinuities and entrapped air. The free end of the piece ofbiophotonic material is then doubled back nearly touching itself so thatthe angle of removal of the piece from the steel plate will be 180degrees. The free end of the piece of biophotonic material (the onepulled) is attached to the adhesion tester scale (an Instron tensiletester or Harvey tensile tester). The test plate is then clamped in thejaws of the tensile testing machine capable of moving the plate awayfrom the scale at a predetermined constant rate. The scale reading in kgis recorded as the tape is peeled from the steel surface.

In some embodiments, the biophotonic material has an adhesion strengththat is less than its tear strength. In some embodiments, thebiophotonic material has an adhesion strength of from about 0.01 N/mm toabout 0.60 N/mm. In some embodiments, the adhesion strength is fromabout 0.20 N/mm to about 0.40 N/mm, or from about 0.25 N/mm to about0.35 N/mm.

In some embodiments, the adhesion strength is less than about 0.10 N/mm,less than about 0.15 N/mm, less than about 0.20 N/mm, less than about0.25 N/mm, less than about 0.30 N/mm, less than about 0.35 N/mm, lessthan about 0.40 N/mm, less than about 0.45 N/mm, less than about 0.55N/mm or less than about 0.60 N/mm.

(6) Methods of Us

The biophotonic materials of the present disclosure may have cosmeticand/or medical benefits. They can be used to promote skin rejuvenationand skin conditioning, promote the treatment of a skin disorder such asacne, promote tissue repair, and promote wound healing includingperiodontitis pockets. They can be used to treat acute inflammation.Acute inflammation can present itself as pain, heat, redness, swellingand loss of function. It includes those seen in allergic reactions suchas insect bites e.g.; mosquito, bees, wasps, poison ivy, post-ablativetreatment.

Accordingly, in certain embodiments, the present disclosure provides amethod for treating acute inflammation.

In certain embodiments, the present disclosure provides a method forproviding skin rejuvenation, treating a skin disorder and/oraccelerating wound healing and/or tissue repair, the method comprising:applying a biophotonic material of the present disclosure to the area ofthe skin or tissue in need of treatment, and illuminating biophotonicmaterial with light having a wavelength that overlaps with an absorptionspectrum of the chromophore(s) present in the biophotonic

In the methods of the present disclosure, any source of actinic lightcan be used. Any type of halogen, LED or plasma arc lamp, or laser maybe suitable. The primary characteristic of suitable sources of actiniclight will be that they emit light in a wavelength (or wavelengths)appropriate for activating the one or more photoactivators present inthe composition. In one embodiment, an argon laser is used. In anotherembodiment, a potassium-titanyl phosphate (KTP) laser (e.g. aGreenLight™ laser) is used. In yet another embodiment, a LED lamp suchas a photocuring device is the source of the actinic light. In yetanother embodiment, the source of the actinic light is a source of lighthaving a wavelength between about 200 to 800 nm. In another embodiment,the source of the actinic light is a source of visible light having awavelength between about 400 and 600 nm. In yet another embodiment, thesource of the actinic light is blue light. In yet another embodiment,the source of the actinic light is red light. In yet another embodiment,the source of the actinic light is green light. Furthermore, the sourceof actinic light should have a suitable power density. Suitable powerdensity for non-collimated light sources (LED, halogen or plasma lamps)are in the range from about 0.1 mW/cm² to about 200 mW/cm². Suitablepower density for laser light sources are in the range from about 0.5mW/cm² to about 0.8 mW/cm².

In some embodiments of the methods of the present disclosure, the lighthas an energy at the subject's skin surface of between about 0.1 mW/cm²and about 500 mW/cm², or 0.1-300 mW/cm², or 0.1-200 mW/cm², wherein theenergy applied depends at least on the condition being treated, thewavelength of the light, the distance of the skin from the light sourceand the thickness of the biophotonic material. In certain embodiments,the light at the subject's skin in between about 1-40 mW/cm², or 20-60mW/cm², or 40-80 mW/cm², or 60-100 mW/cm², or 80-120 mW/cm², or 100-140mW/cm², or 30-180 mW/cm², or 120-160 mW/cm², or 140-180 mW/cm², or160-200 mW/cm², or 110-240 mW/cm², or 110-150 mW/cm², or 190-240 mW/cm².

The activation of the photoactivators within the biophotonic materialmay take place almost immediately on illumination (femto- or picaseconds). A prolonged exposure period may be beneficial to exploit thesynergistic effects of the absorbed, reflected and reemitted light ofthe biophotonic material of the present disclosure and its interactionwith the tissue being treated. In one embodiment, the time of exposureto actinic light of the tissue or skin or biophotonic material is aperiod between 1 minute and 5 minutes. In another embodiment, the timeof exposure to actinic light of the tissue or skin or biophotonicmaterial is a period between 1 minute and 5 minutes. In some otherembodiments, the biophotonic material is illuminated for a periodbetween 1 minute and 3 minutes. In certain embodiments, light is appliedfor a period of 1-30 seconds, 15-45 seconds, 30-60 seconds, 0.75-1.5minutes, 1-2 minutes, 1.5-2.5 minutes, 2-3 minutes, 2.5-3.5 minutes, 3-4minutes, 3.5-4.5 minutes, 4-5 minutes, 5-10 minutes, 10-15 minutes,15-20 minutes, or 20-30 minutes. In certain embodiments, the biophotonicmaterial may be re-illuminated at certain intervals. In yet anotherembodiment, the source of actinic light is in continuous motion over thetreated area for the appropriate time of exposure.

In certain embodiments, the first and/or the second chromophore (whenpresent) in the cohesive matrix can be photoexcited by ambient lightincluding from the sun and overhead lighting. In certain embodiments,the first and/or the second chromophore (when present) can bephotoactivated by light in the visible range of the electromagneticspectrum. The light can be emitted by any light source such as sunlight,light bulb, an LED device, electronic display screens such as on atelevision, computer, telephone, mobile device, flashlights on mobiledevices. In the methods of the present disclosure, any source of lightcan be used. For example, a combination of ambient light and directsunlight or direct artificial light may be used. Ambient light caninclude overhead lighting such as LED bulbs, fluorescent bulbs etc, andindirect sunlight.

In the methods of the present disclosure, the biophotonic material maybe removed from the skin following application of light. In someembodiments the biophotonic material is peeled off from the skinfollowing application of light. In some embodiments, the biophotonicmaterial is removed as a single piece from the skin followingapplication of light. In other embodiments, the biophotonic material isleft on the tissue for an extended period of time and re-activated withdirect or ambient light at appropriate times to treat the condition.

In certain embodiments of the method of the present disclosure, thebiophotonrc material can be applied to the tissue, such as on the face,once, twice, three times, four times, five times or six times a week,daily, or at any other frequency. The total treatment time can be oneweek, two weeks, three weeks, four weeks, five weeks, six weeks, sevenweeks, eight weeks, nine weeks, ten weeks, eleven weeks, twelve weeks,or any other length of time deemed appropriate. In certain embodiments,the total tissue area to be treated may be split into separate areas(cheeks, forehead), and each area treated separately. For example, thecomposition may be applied topically to a first portion, and thatportion illuminated with light, and the biophotonic composition thenremoved. Then the composition is applied to a second portion,illuminated and removed. Finally, the composition is applied to a thirdportion, illuminated and removed.

In certain embodiments, the biophotonic material can be used followingwound closure to optimize scar revision. In this case biophotonicmaterial may be applied at regular intervals such as once a week, or atan interval deemed appropriate by the physician.

In certain embodiments, the biophotonic material can be used followingacne treatment to maintain the condition of the treated skin. In thiscase, the biophotonic material may be applied at regular intervals suchas once a week, or at an interval deemed appropriate by the physician.

In certain embodiments, the biophotonic material can be used followingablative skin rejuvenation treatment to maintain the condition of thetreated skin. In this case, the biophotonic material may be applied atregular intervals such as once a week, or at an interval deemedappropriate by the physician.

In the methods of the present disclosure, additional components mayoptionally be included in the biophotonic materials or used incombination with the biophotonic materials. Such additional componentsinclude, but are not limited to, healing factors, antimicrobials,oxygen-rich agents, wrinkle fillers such as botox, hyaluronic acid andpolylactic acid, fungal, anti-bacterial, anti-viral agents and/or agentsthat promote collagen synthesis. These additional components may beapplied to the skin in a topical fashion, prior to, at the same time of,and/or after topical application of the biophotonic materials of thepresent disclosure. Suitable healing factors comprise compounds thatpromote or enhance the healing or regenerative process of the tissues onthe application site. During the photoactivation of a biophotonicmaterial of the present disclosure, there may be an increase of theabsorption of molecules of such additional components at the treatmentsite by the skin or the mucosa. In certain embodiments, an augmentationin the blood flow at the site of treatment can observed for a period oftime. An increase in the lymphatic drainage and a possible change in theosmotic equilibrium due to the dynamic interaction of the free radicalcascades can be enhanced or even fortified with the inclusion of healingfactors. Suitable healing factors include, but are not limited toglucosamines, allantoins, saffron, agents that promote collagensynthesis, anti-fungal, anti-bacterial, anti-viral agents and woundhealing factors such as growth factors.

(i) Skin Rejuvenation

The biophotonic material of the present disclosure may be useful inpromoting skin rejuvenation. The dermis is the second layer of skin,containing the structural elements of the skin, the connective tissue.There are various types of connective tissue with different functions.Elastin fibers give the skin its elasticity, and collagen gives the skinits strength.

The junction between the dermis and the epidermis is an importantstructure. The dermal-epidermal junction interlocks forming finger-likeepidermal ridges. The cells of the epidermis receive their nutrientsfrom the blood vessels in the dermis. The epidermal ridges increase thesurface area of the epidermis that is exposed to these blood vessels andthe needed nutrients.

The aging of skin comes with significant physiological changes to theskin. The generation of new skin cells slows down, and the epidermalridges of the dermal-epidermal junction flatten out. While the number ofelastin fibers increases, their structure and coherence decreases. Alsothe amount of collagen and the thickness of the dermis decrease with theageing of the skin.

Collagen is a major component of the skin's extracellular matrix,providing a structural framework. During the aging process, the decreaseof collagen synthesis and insolubilization of collagen fibers contributeto a thinning of the dermis and loss of the skin's biomechanicalproperties.

The physiological changes to the skin result in noticeable agingsymptoms often referred to as chronological-, intrinsic- andphoto-ageing. The skin becomes drier, roughness and scaling increase,the appearance becomes duller, and most obviously fine lines andwrinkles appear. Other symptoms or signs of skin aging include, but arenot limited to, thinning and transparent skin, loss of underlying fat(leading to hollowed cheeks and eye sockets as well as noticeable lossof firmness on the hands and neck), bone loss (such that bones shrinkaway from the skin due to bone loss, which causes sagging skin), dryskin (which might itch), inability to sweat sufficiently to cool theskin, unwanted facial hair, freckles, age spots, spider veins, rough andleathery skin, fine wrinkles that disappear when stretched, loose skin,a blotchy complexion.

The dermal-epidermal junction is a basement membrane that separates thekeratinocytes in the epidermis from the extracellular matrix, which liesbelow in the dermis. This membrane consists of two layers: the basallamina in contact with the keratinocytes, and the underlying reticularlamina in contact with the extracellular matrix. The basal lamina isrich in collagen type IV and laminin, molecules that play a role inproviding a structural network and bioadhesive properties for cellattachment.

Laminin is a glycoprotein that only exists in basement membranes. It iscomposed of three polypeptide chains (alpha, beta and gamma) arranged inthe shape of an asymmetric cross and held together by disulfide bonds.The three chains exist as different subtypes which result in twelvedifferent isoforms for laminin, including Laminin-1 and Laminin-5.

The dermis is anchored to hemidesmosomes, specific junction pointslocated on the keratinocytes, which consist of α-integrins and otherproteins, at the basal membrane keratinocytes by type VII collagenfibrils. Laminins, and particularly Laminin-5, constitute the realanchor point between hemidesmosomal transmembrane proteins in basalkeratinocytes and type VII collagen.

Laminin-5 synthesis and type VII collagen expression have been proven todecrease in aged skin. This causes a loss of contact between dermis andepidermis, and results in the skin losing elasticity and becoming saggy.

Recently another type of wrinkles, generally referred to as expressionwrinkles, got general recognition. These wrinkles require loss ofresilience, particularly in the dermis, because of which the skin is nolonger able to resume its original state when facial muscles whichproduce facial expressions exert stress on the skin, resulting inexpression wrinkles.

The biophotonic material of the present disclosure and methods of thepresent disclosure promote skin rejuvenation. In certain embodiments,the biophotonic material and methods of the present disclosure promotecollagen synthesis. In certain other embodiments, the biophotonicmaterial and methods of the present disclosure may reduce, diminish,retard or even reverse one or more signs of skin aging including, butnot limited to, appearance of fine lines or wrinkles, thin andtransparent skin, loss of underlying fat (leading to hollowed cheeks andeye sockets as well as noticeable loss of firmness on the hands andneck), bone loss (such that bones shrink away from the skin due to boneloss, which causes sagging skin), dry skin (which might itch), inabilityto sweat sufficiently to cool the skin, unwanted facial hair, freckles,age spots, spider veins, rough and leathery skin, fine wrinkles thatdisappear when stretched, loose skin, or a blotchy complexion. Incertain embodiments, the biphotonic material and methods of the presentdisclosure may induce a reduction in pore size, enhance sculpturing ofskin subsections, and/or enhance skin translucence.

In certain embodiments, the biophotonic material may be used inconjunction with collagen promoting agents. Agents that promote collagensynthesis (i.e., pro-collagen synthesis agents) include amino acids,peptides, proteins, lipids, small chemical molecules, natural productsand extracts from natural products.

For instance, it was discovered that intake of vitamin C, iron, andcollagen can effectively increase the amount of collagen in skin orbone. See, e.g., U.S. Patent Application Publication 20090069217.Examples of the vitamin C include an ascorbic acid derivative such asL-ascorbic acid or sodium L-ascorbate, an ascorbic acid preparationobtained by coating ascorbic acid with an emulsifier or the like, and amixture containing two or more of those vitamin Cs at an arbitrary rate.In addition, natural products containing vitamin C such as acerola andlemon may also be used. Examples of the iron preparation include: aninorganic iron such as ferrous sulfate, sodium ferrous citrate, orferric pyrophosphate; an organic iron such as heme iron, ferritin iron,or lactoferrin iron; and a mixture containing two or more of those ironsat an arbitrary rate. In addition, natural products containing iron suchas spinach or liver may also be used. Moreover, examples of the collageninclude: an extract obtained by treating bone, skin, or the like of amammal such as bovine or swine with an acid or alkaline; a peptideobtained by hydrolyzing the extract with a protease such as pepsin,trypsin, or chymotrypsin; and a mixture containing two or more of thosecollagens at an arbitrary rate. Collagens extracted from plant sourcesmay also be used.

Additional pro-collagen synthesis agents are described, for example, inU.S. Pat. No. 7,598,291, 7,722,904, 6,203,805, 5,529,769, etc, and U.S.Patent Application Publications 20060247313, 20080108681, 20110130459,20090325885, 20110086060, etc.

(ii) Skin disorders

The biophotonic materials and methods of the present disclosure may beused to treat skin disorders that include, but are not limited to,erythema, telangiectasia, actinic telangiectasia, psoriasis, skincancer, pemphigus, sunburn, dermatitis, eczema, rashes, impetigo, lichensimplex chronicus, rhinophyma, perioral dermatitis, pseudofolliculitisbarbae, drug eruptions, erythema multiforme, erythema nodosum,granuloma. annulare, actinic keratosis, purpura, alopecia areata,aphthous stomatitis, drug eruptions, dry skin, chapping, xerosis,ichthyosis vulgaris, fungal infections, herpes simplex, intertrigo,keloids, keratoses, milia, moluscum contagiosum, pityriasis rosea,pruritus, urticaria, and vascular tumors and malformations. Dermatitisincludes contact dermatitis, atopic dermatitis, seborrheic dermatitis,nummular dermatitis, generalized exfoliative dermatitis, and statisdermatitis. Skin cancers include melanoma, basal cell carcinoma, andsquamous cell carcinoma.

(iii) Acne and Acne Scars

The biophotonic materials and methods of the present disclosure may beused to treat acne. As used herein, “acne” means a disorder of the skincaused by inflammation of skin glands or hair follicles. The biophotonicmaterials and methods of the disclosure can be used to treat acne atearly pre-emergent stages or later stages where lesions from acne arevisible. Mild, moderate and severe acne can be treated with embodimentsof the biophotonic compositions and methods. Early pre-emergent stagesof acne usually begin with an excessive secretion of sebum or dermal oilfrom the sebaceous glands located in the pilosebaceous apparatus. Sebumreaches the skin surface through the duct of the hair follicle. Thepresence of excessive amounts of sebum in the duct and on the skin tendsto obstruct or stagnate the normal flow of sebum from the follicularduct, thus producing a thickening and solidification of the sebum tocreate a solid plug known as a comedone. In the normal sequence ofdeveloping acne, hyperkeratinazation of the follicular opening isstimulated, thus completing blocking of the duct. The usual results arepapules, pustules, or cysts, often contaminated with bacteria, whichcause secondary infections. Acne is characterized particularly by thepresence of comedones, inflammatory papules, or cysts. The appearance ofacne may range from slight skin irritation to pitting and even thedevelopment of disfiguring scars. Accordingly, the biophotonic materialsand methods of the present disclosure can be used to treat one or moreof skin irritation, pitting, development of scars, comedones,inflammatory papules, cysts, hyperkeratinazation, and thickening andhardening of sebum associated with acne.

Some types of acne include, for example, acne vulgaris, cystic acne,acne atrophica, bromide acne, chlorine acne, acne conglobata, acne cosetica, acne detergicans, epidemic acne, acne estivalis, acne fulminans,halogen acne, acne indurata, iodide acne, acne keloid, acne mechanica,acne papulosa, pomade acne, premenstal acne, acne pustulosa, acnescorbutica, acne scrofulosorum, acne urticata, acne varioliformis, acnevenenata, propionic acne, acne excoriee, gram negative acne, steroidacne, and nodulocystic acne. Some skin disorders present varioussymptoms including redness, flushing, burning, scaling, pimples,papules, pustules, comedones, macules, nodules, vesicles, blisters,telangiectasia, spider veins, sores, surface irritations or pain,itching, inflammation, red, purple, or blue patches or discolorations,moles, and/or tumors.

The biophotonic materials and methods of the present disclosure may beused to treat various types of acne. Some types of acne include, forexample, acne vulgaris, cystic acne, acne atrophica, bromide acne,chlorine acne, acne conglobata, acne cosmetica, acne detergicans,epidemic acne, acne estivalis, acne fulminans, halogen acne, acneindurata, iodide acne, acne keloid, acne mechanica, acne papulosa,pomade acne, premenstral acne, acne pustulosa, acne scorbutica, acnescrofulosorum, acne urticata, acne varioliformis, acne venenata,propionic acne, acne excoriee, gram negative acne, steroid acne, andnodulocystic acne.

In certain embodiments, the biophotonic material of the presentdisclosure is used in conjunction with systemic or topical antibiotictreatment. For example, antibiotics used to treat acne includetetracycline, erythromycin, minocycline, doxycycline, which may also beused with the compositions and methods of the present disclosure. Theuse of the biophotonic material can reduce the time needed for theantibiotic treatment or reduce the dosage.

(iv) Wound Healing

The biophotonic materials and methods of the present disclosure may beused to treat wounds, promote wound healing or promote tissue repair.Wounds that may be treated by the biophotonic materials and methods ofthe present disclosure include, for example, injuries to the skin andsubcutaneous tissue initiated in different ways (e.g., pressure ulcersfrom extended bed rest, wounds induced by trauma, wounds induced byconditions such as periodontitis) and with varying characteristics. Incertain embodiments, the present disclosure provides biophotonicmaterials and methods for treating and/or promoting the healing of, forexample, burns, incisions, excisions, lesions, lacerations, abrasions,puncture or penetrating wounds, surgical wounds, contusions, hematomas,crushing injuries, amputations, sores and ulcers.

Biophotonic materials and methods of the present disclosure may be usedto treat and/or promote the healing of chronic cutaneous ulcers orwounds, which are wounds that have failed to proceed through an orderlyand timely series of events to produce a durable structural, functional,and cosmetic closure. The vast majority of chronic wounds can beclassified into three categories based on their etiology: pressureulcers, neuropathic (diabetic foot) ulcers and vascular (venous orarterial ) ulcers.

For example, the present disclosure provides biophotonic materials andmethods for treating and/or promoting healing of a diabetic ulcer.Diabetic patients are prone to foot and other ulcerations due to bothneurologic and vascular complications. Peripheral neuropathy can causealtered or complete loss of sensation in the foot and/or leg. Diabeticpatients with advanced neuropathy lose all ability for sharp-dulldiscrimination. Any cuts or trauma to the foot may go completelyunnoticed for days or weeks in a patient neuropathy. A patient withadvanced neuropathy loses the ability to sense a sustained pressureinsult, as a result, tissue ischemia and necrosis may occur leading tofor example, plantar ulcerations. Microvascular disease is one of thesignificant complications for diabetics which may also lead toulcerations. In certain embodiments, biophotonic materials and methodsof treating a chronic wound are provided here in, where the chronicwound is characterized by diabetic foot ulcers and/or ulcerations due toneurologic and/or vascular complications of diabetes.

In other examples, the present disclosure provides biophotonic materialsand methods for treating and/or promoting healing of a pressure ulcer.Pressure ulcers include bed sores, decubitus ulcers and ischialtuberosity ulcers and can cause considerable pain and discomfort to apatient. A pressure ulcer can occur as a result of a prolonged pressureapplied to the skin. Thus, pressure can be exerted on the skin of apatient due to the weight or mass of an individual. A pressure ulcer candevelop when blood supply to an area of the skin is obstructed or cutoff for more than two or three hours. The affected skin area can turnred, become painful and necrotic. If untreated, the skin can break openand become infected. A pressure ulcer is therefore a skin ulcer thatoccurs in an area of the skin that is under pressure from e.g. lying inbed, sitting in a wheelchair, and/or wearing a cast for a prolongedperiod of time. Pressure ulcers can occur when a person is bedridden,unconscious, unable to sense pain, or immobile. Pressure ulcers oftenoccur in boney prominences of the body such as the buttocks area (on thesacrum or iliac crest), or on the heels of foot.

Additional types of wounds that ca.n be treated by the biophotonicmaterials and methods of the present disclosure include those disclosedby U.S. Pat. Appl. Publ. No. 20090220450, which is incorporated hereinby reference.

There are three distinct phases in the wound healing process. First, inthe inflammatory phase, which typically occurs from the moment a woundoccurs until the first two to five days, platelets aggregate to depositgranules, promoting the deposit of fibrin and stimulating the release ofgrowth factors. Leukocytes migrate to the wound site and begin to digestand transport debris away from the wound. During this inflammatoryphase, monocytes are also converted to macrophages, which release growthfactors for stimulating angiogenesis and the production of fibroblasts.

Second, in the proliferative phase, which typically occurs from two daysto three weeks, granulation tissue forms, and epithelialization andcontraction begin. Fibroblasts, which are key cell types in this phase,proliferate and synthesize collagen to fill the wound and provide astrong matrix on which epithelial cells grow. As fibroblasts producecollagen, vascularization extends from nearby vessels, resulting ingranulation tissue. Granulation tissue typically grows from the base ofthe wound. Epithelialization involves the migration of epithelial cellsfrom the wound surfaces to seal the wound. Epithelial cells are drivenby the need to contact cells of like type and are guided by a network offibrin strands that function as a grid over which these cells migrate.Contractile cells called myofibroblasts appear in wounds, and aid inwound closure. These cells exhibit collagen synthesis and contractility,and are common in granulating wounds.

Third, in the remodeling phase, the final phase of wound healing whichcan take place from three weeks up to several years, collagen in thescar undergoes repeated degradation and re-synthesis. During this phase,the tensile strength of the newly formed skin increases.

However, as the rate of wound healing increases, there is often anassociated increase in scar formation. Scarring is a consequence of thehealing process in most adult animal and human tissues. Scar tissue isnot identical to the tissue which it replaces, as it is usually ofinferior functional quality. The types of scars include, but are notlimited to, atrophic, hypertrophic and keloidal scars, as well as scarcontractures. Atrophic scars are and depressed below the surroundingskin as a valley or hole. Hypertrophic scars are elevated scars thatremain within the boundaries of the original lesion, and often containexcessive collagen arranged in an abnormal pattern. Keloidal scars areelevated scars that spread beyond the margins of the original wound andinvade the surrounding normal skin in a way that is site specific, andoften contain whorls of collagen arranged in an abnormal fashion.

In contrast, normal skin consists of collagen fibers arranged in abasket-weave pattern, which contributes to both the strength andelasticity of the dermis. Thus, to achieve a smoother wound healingprocess, an approach is needed that not only stimulates collagenproduction, but also does so in a way that reduces scar formation.

The biophotonic materials and methods of the present disclosure promotethe wound healing by promoting the formation of substantially uniformepithelialization; promoting collagen synthesis; promoting controlledcontraction; and/or by reducing the formation of scar tissue. In certainembodiments, the biophotonic materials and methods of the presentdisclosure may promote wound healing by promoting the formation ofsubstantially uniform epithelialization. In some embodiments, thebiophotonic materials and methods of the present disclosure promotecollagen synthesis. In some other embodiments, the biophotonic materialsand methods of the present disclosure promote controlled contraction. Incertain embodiments, the biophotonic erials and methods of the presentdisclosure promote wound healing, for example, by reducing the formationof scar tissue.

In the methods of the present disclosure, the biophotonic materials ofthe present disclosure may also be used in combination with negativepressure assisted would closure devices and systems.

In certain embodiments, the biophotonic material is kept in place for upto one, two or 3 weeks, and illuminated with light which may includeambient light at various intervals. In this case, the composition may becovered up in between exposure to light with an opaque material or leftexposed to light.

(6) Kits

The present disclosure also provides kits for preparing a biophotonicmaterial and/or providing any of the components required for formingbiophotonic materials of the present disclosure.

In some embodiments, the kit includes containers comprising thecomponents or compositions that can be used to make the biophotonicmaterials of the present disclosure. In some embodiments, the kitincludes a biophotonic material of the present disclosure. The differentcomponents making up the biophotonic materials of the present disclosuremay be provided in separate containers. For example, if the biophotonicmaterial is to include an oxygen-rich agent, the oxygen-rich agentshould be provided in a container separate from the chromophore.Examples of such containers are dual chamber syringes, or sachets withpouches. Another example is one of the components being provided in asyringe which can be injected into a container of another component.

In other embodiments, the kit comprises a systemic drug for augmentingthe treatment of the biophotonic material of the present disclosure. Forexample, the kit may include a systemic or topical antibiotic, hormonetreatment (e.g. for acne treatment or wound healing), or a negativepressure device.

In certain embodiments, the kit comprises a first component comprising afirst chromophore; and a second component comprising a at least onethickening agent, wherein the thickening agent can form a cohesivematrix when mixed with the first component, when the mixture is appliedto skin, or when illuminated with light.

In other embodiments, the kit comprises a means for applying thecomponents of the biophotonic materials.

In certain aspects, there is provided a container comprising a chamberfor holding a biophotonic material, and an outlet in communication withthe chamber for discharging the biophotonic material from the container,wherein the biophotonic material comprises at least one chromophore in acarrier medium which can form a biophotonic material after beingdischarged from the sealed chamber. The container can be a pressurizedor non-pressurized spray can.

In certain embodiments, the kit comprises a first component comprisingthe biophotonic material or a non-cohesive form of the biophotonicmaterial (‘precursor’), and the second component comprises a dressing ora mask. The dressing or mask may be a porous or semi-porous structurefor receiving the biophotonic material. The dressing or mask may alsocomprise woven or non-woven fibrous materials. The biophotonic materialor its precursor can be incorporated, such as by injection, into thedressing before the biophotonic material takes on a cohesive form withinthe dressing or mask.

In certain embodiments of the kit, the kit may further comprise a lightsource such as a portable light with a wavelength appropriate toactivate the chromophore the biophotonic material. The portable lightmay be battery operated or re-chargeable.

Written instructions on how to use the biophotonic materials inaccordance with the present disclosure may be included in the kit, ormay be included on or associated with the containers comprising thecompositions or components making up the biophotonic materials of thepresent disclosure. The instructions can include information on how toform the cohesive matrix from the thickening agent(s) or matrixprecursors provided with the kit.

Identification of equivalent biophotonic materials, methods and kits arewell within the skill of the ordinary practitioner and would require nomore than routine experimentation, in light of the teachings of thepresent disclosure. Practice of the disclosure will be still more fullyunderstood from the following examples, which are presented herein forillustration only and should not be construed as limiting the disclosurein any way.

EXAMPLES Example 1 Preparation of an exemplary cohesive biophotonicmaterial

% in Composition (wt/wt) Water 60-95 Glycerine  5-15 Propylene Glycol2-6 Sodium hyaluronate 2-8 Urea peroxide 1-5 Glucosamine sulfate 0.5-4 Carbopol 0.1-2  First Chromophore 0.001-0.01  Second chromophore0.001-0.01 

Phase A was prepared by mixing water, Eosin Y, Rose Bengal andGlucosamine sulphate. Phase B (Water, Glycerin, Propylene glycol, Ureaperoxide, Carbopol polymer) was then added to Phase A, and mixed until alight viscous liquid was obtained. Phase C (Sodium hyaluronate) was thenadded to the mixture, and mixed until a homogenous thick gel wasobtained.

The homogenous gel obtained above was spread onto a flat surface. Thelayer was covered with an aluminum sheet and was allowed to dry for 24hours. After 24 hours, the resulting material was a cohesive gel, waselastic and easy to manipulate. It could be applied to the skin andpealed off with little or no residue remaining. A 5-20% weight loss ofthe total weight of the material was found to occur after drying for 24hours. The film could be stored between two layers of saran wrap,paraffin etc. On illumination with light (peak wavelength between400-470nm and a power density of about 30-150 mW/cm²) for 5 minutes at adistance of 5 cm from the light source, the film emitted fluorescentlight which was captured by a photospectrometer and is illustrated inFIG. 3. The emitted fluorescent light was in the green, yellow andorange portions of the electromagnetic spectrum.

Example 2 Measurement of Tensile Strength

The tensile strength of a biophotonic material formed according toExample 1 may be measured by the following method. Pieces of thebiophotonic material having a size of about 2 cm×2 cm, and 2 mmthickness are prepared. The tensile strength of the biophotonic materialpieces is then measured using a 500N capacity tabletop mechanicaltesting system (#5942R4910, Instron) with a 5N maximum static load cell(#102608, Instron). Pneumatic side action grips are used to secure thesamples (#2712-019, Instron). A constant extension rate of 2 mm/minuntil failure is used and the tensile strength is calculated from thestress vs. strain data plots. Stress-strain testing is detailed bystandards-setting organizations, notably the American Society forTesting and Materials (ASTM). A preferred method is ASTM D638.

Example 3 Measurement of Tear Strength

The tear strength of a biophotonic material formed according to Example1 may be measured by any American Society for Testing and Materials(ASTM) methods known in the art. A preferred method is ASTM D1004.

Example 4 Measurement of Adhesion Strength

The adhesion strength of the biophotonic material prepared according toExample 1 is measured in Newtons per meter (N/m) using a peel test withan Instron 1122 Tensile Tester. First, pieces of the biophotonicmaterial having a size of about 2 cm×2 cm, and 2. mm thickness areprepared, placed onto a glass plate and then then mounted into theInstron 1122 Tensile Tester with the substrate clamped in the upper gripand the glass plate clamped in the bottom grip. The average force (N)required to peel the piece of biophotonic solid off the glass plate at a180° angle at speed of 50 min/min is recorded. Using this force valuethe adhesive strength in units of N/m is calculated using the equation:

S _(A) =F _(P)(1−cos θ)/w

wherein S_(A) is the adhesive strength, F_(P) is the peel force, θ isthe angle of peel (180°), and w is the width of the sample (2 cm).

Example 5 Angiogenic Potential of the Biophotonic Material of theDisclosure

A human skin model was developed to assess the angiogenic potential ofthe biophotonic material of the present disclosure. Briefly, abiophotonic composition comprising a fluorescent chromophore was placedon top of a human skin model containing fibroblasts and keratinocytes.The skin model and the composition were separated by a nylon mesh of 20micron pore size. The composition was then irradiated with blue light(‘activating light’) for 5 minutes at a distance of 5 cm from the lightsource. The activating light consisted of light emitted from an LED lamphaving an average peak wavelength of about 400-470 nm, and a powerintensity measured at 10 cm of 7.7 J/cm² to 11.5 J/cm². Uponillumination with the activating light, the biophotonic compositionemitted fluorescent light (FIG. 4) which may be replicated by abiophotonic material of the present disclosure. Since the biophotoniccomposition was in limited contact with the cells, the fibroblasts andkeratinocytes were exposed mainly to the activating light and thefluorescent light emitted from the biophotonic composition. Conditionedmedia from the treated human 3D skin model were then applied to humanaortic endothelial cells previously plated in matrigel. The formation oftubes by endothelial cells was observed and monitored by microscopyafter 24 hours, The conditioned medium from 3D skin models treated withlight illumination induced endothelial tube formation in vitro,suggesting an indirect effect of the light treatment (blue light andfluorescence) on angiogenesis via the production of factors byfibroblasts and keratinocytes. Plain medium and conditioned medium ofuntreated skin samples were used as a control, and did not induceendothelial tube formation.

Example 6 Protein Secretion and Gene Expression Profiles

Wounded and unwounded 3D human skin models (EpiDermFT, MatTekCorporation) were used assess the potential of a biophotonic material ofthe present disclosure to trigger distinct protein secretion and geneexpression profiles. Briefly, a biophotonic composition comprising Eosinand Erythrosine were placed on top of wounded and unwounded 3D humanskin models cultured under different conditions (with growth factors,50% growth factors and no growth factors). The skin models and thecomposition were separated by a nylon mesh of 20 micron pore size. Eachskin model-composition combination was then irradiated with blue light(‘activating light’) for 5 minutes at a distance of 5 cm from the lightsource. The activating light consisted of light emitted from an LED lamphaving an average peak wavelength of about 440-470 nm, a power densityof 60-150 mW/cm2 at 5 cm, and a total intensity after 5 minutes of about18-39 J/cm2. The controls were consisted of 3D skin models notilluminated with light.

Gene expression and protein secretion profiles were measured 24 hourspost-light exposure. Cytokine secretion was analyzed by antibody arrays(RayBio Human Cytokine antibody array), gene expression was analyzed byPCR array (PAHS-013A, SABioscience) and cytotoxicity was determined byGAPDH and LDH release. Results (Tables 1 and 2) showed that the lighttreatment is capable of increasing the level of protein secreted andgene expression involved in the early inflammatory phase of woundhealing in wounded skin inserts and in non-starvation conditions. Instarvation conditions mimicking chronic wounds, there was no increase inthe level of inflammatory protein secreted when compared to the control.Interestingly, the effect of the light treatment on unwounded skinmodels has a much lower impact at the cellular level than on woundedskin insert, which suggests an effect at the cellular effect level ofthe light treatment. It seems to accelerate the inflammatory phase ofthe wound healing process. Due to the lack of other cell types such asmacrophages in the 3D skin model, the anti-inflammatory feed-back isabsent and may explain the delay in wound closure. Cytoxicity was notobserved in the light treatments.

TABLE 1 List of proteins with statistically significant differencesecretion ratio between treated and untreated control at day 3. Medium1X Medium 0.5X Medium 0X Increase ENA78 p = 0.04 ↑↑ Angiogenin p = 0.03↑ II-1R4/ST2 p = 0.02 ↑↑ CXCL16 p = 0.04 ↑ MMP3 p = 0.01 ↑↑ MCP-2 p =0.04 ↑↑ Decrease BMP6 p = 0.01 ↓ BMP6 p = 0.02 ↓ TNFα p = 0.005 ↓ Twoarrows mean that the ratio was over 2 folds.

TABLE 2 List of genes with statistically significant differenceexpression ratio between treated and untreated control during the first24 hours. Medium 1X Medium 0.5X Medium 0X Increase CTGF p = 0.02 ↑ CTGFP = 0.04 ↑ MMP3 p = 0.007 ↑↑ ITGB3 p = 0.03 ↑ ITGB3 p = 0.05 ↑ LAMA1 p =0.03 ↑ MMP1 p = 0.03 ↑ MMP1 p = 0.02 ↑↑ ITGA2 p = 0.03 ↑ MMP3 p = 0.01 ↑MMP10 p = 0.003 ↑↑ THBS1 P = 0.02 ↑ MMP3 p = 0.007 ↑↑ MMP8 p = 0.02 ↑↑THBS1 p = 0.03 ↑ Decrease HAS1 p = 0.009 ↓↓ NCAM1 p = 0.02 ↓↓ NCAM1 p =0.05 ↓↓ VCAN p = 0.02 ↓ VCAM1 p = 0.03 ↓↓ LAMC1 p = 0.002 ↓ COL7A1 p =0.04 ↓ COL6A1 p = 0.007 ↓ CTNNA1 p = 0.03 ↓ MMP7 p = 0.003 ↓ Two arrowsmean that the ratio was over 2 folds.

Example 7 Selecting Concentration of Chromophore in Composition

The fluorescence spectra of biophotonic materials with differentconcentrations of chromophores were investigated using aspectrophotometer and an activating blue light. Exemplary fluorescencespectra of Eosin Y and Fluorescein are presented in FIGS. 5a and 5 b,respectively. It was found that emitted fluorescence from thechromophore increases rapidly with increasing concentration but slowsdown to a plateau with further concentration increase. Activating lightpassing through the composition decreases with increasing chromophorecomposition as more is absorbed by the chromophores. Therefore, theconcentration of chromophores in biophotonic materials of the presentdisclosure can be selected according to a required ratio and level ofactivating light and fluorescence treating the tissue based on thisexample.

Example 8 Synergistic Combination of Eosin Y and Fluorescein

The photodynamic properties of (i) Fluorescein sodium salt at about 0.09mg/mL, (ii) Eosin Y at about 0.305 mg/mL, and (iii) a mixture ofFluorescein sodium salt at about 0.09 mg/mL and Eosin Y at about 0.305mg/mL in a gel (comprising about 12% carbamide peroxide), wereevaluated. A flexstation 384 II spectrometer was used with the followingparameters: mode fluorescence, excitation 460 nm, emission spectra465-750 nm. The absorption and emission spectra are shown in FIGS. 6aand 6b which indicate an energy transfer between the chromophores in thecombination. It is to be reasonably inferred that this energy transfercan also occur in biophotonic materials of the present disclosure.

Example 9 Synergistic Combination of Eosin y, Fluorescein and RoseBengal

The photodynamic properties of (i) Rose Bengal at about 0.085 mg/mL,(ii) Fluorescein sodium salt at about 0.44 mg/mL final concentration,(ii) Eosin Y at about 0.305 mg/mL, and (iii) a mixture of (i), (ii) and(iii) in a gel (comprising about 12% carbamide peroxide) (Set A), wereevaluated. A flexstation 384 II spectrometer was used with the followingparameters: mode fluorescence, excitation 460 nm, emission spectra465-750 nm. The absorbance and emission spectra are shown in FIGS. 7aand 7b which indicate an energy transfer between the chromophores in thechromophore combination. It is to be reasonably inferred that thisenergy transfer can also occur in biophotonic materials of the presentdisclosure.

Energy transfer was also seen between: Eosin Y and Rose Bengal; PhloxineB and EosinY; Phloxine B, EosinY and Fluorescein, amongst othercombinations. It is to be reasonably inferred that energy transfer canalso occur in biophotonic materials of the present disclosure.

Example 10 Eosin Y and Fluorescein Induce Collagen Formation

A composition according to an embodiment of the present invention,comprising 0.01% Eosin Y and 0.01% Fluorescein in a carrier matrix (1.8%carbopol gel) was evaluated for its potential to induce collagenformation. Dermal human fibroblasts were plated in glass-bottomed disheswith wells (MatTek®). There were approximately 4000 cells per well.After 48 hours, the glass-bottomed dishes were inverted and the cellswere treated through the glass bottom with (i) a no light (control),(ii) sunlight exposure for about 13 minutes at noon (control), (iii) thecomposition applied to the glass well bottom on the other side of thecells (no light exposure), and (iv) the composition applied to the glasswell bottom on the other side of the cells (sun light exposure for about13 minutes at noon). In the case of (iii) and (iv), there was no directcontact between the cells and the composition. In the case of (iv), thecells were exposed to emitted light from and through the Eosin Y andFluorescein composition when exposed to sunlight. A partialphotobleaching was observed in (iv). After the treatment, the cells werewashed and incubated in regular medium for 48 hours. A collagen assaywas then performed on the supernatant using the Picro-Sirius red method.This involved adding Sirius red dye solution in picric acid to thesupernatant, incubating with gentle agitation for 30 minutes followed bycentrifugation to form a pellet. The pellet was washed first with 0.1NHCl and then 0.5 N NaOH to remove free dye. After centrifugation, thesuspension was read at 540 nm for collagen type I. The results are shownin Table 3.

TABLE 3 A qualitative comparison of collagen type I concentration in adermal human fibroblast supernatant exposed to (i) a no light (control),(ii) sunlight exposure for about 13 minutes at noon (control), (iii) anylight emitted from a Eosin Y and Fluorescein composition through a glassseparation (no light exposure), and (iv) any light emitted from a EosinY and Fluorescein composition through a glass separation (sun lightexposure for about 13 minutes at noon). Eosin Y and Eosin and No lightSunlight alone Fluorescein - Fluorescein (control) (alone) no lightsunlight Collagen + + ++ +++ formation ++ indicates collagen levelsabout twice as high as +, and +++ indicates collagen levels about threetimes as high as +.

There was a statistical difference between the collagen levels inducedby the Eosin Y and Fluorescein composition exposed to sunlight comparedto the no light and sunlight alone controls.

Collagen generation is indicative of a potential for tissue repairincluding stabilization of granulation tissue and decreasing of woundsize. It is also linked to reduction of fine lines, a decrease in poresize, improvement of texture and improvement of tensile strength ofintact skin.

What is claimed is:
 1. A biophotonic material, said biophotonic materialcomprising: a cohesive matrix, and at least one chromophore, wherein theat least one chromophore can absorb and emit light from within thebiophotonic material.
 2. The biophotonic material of claim 1, whereinthe biophotonic material is elastic.
 3. The biophotonic material ofclaim 2, wherein the biophotonic material is a peelable film.
 4. Thebiophotonic material of claim 1, wherein the tear and/or tensilestrength of the biophotonic material is greater than an adhesivestrength of the biophotonic material to a surface to which it isapplied.
 5. The biophotonic material of claims 1, wherein thebiophotonic material is non-elastic.
 6. The biophotonic material ofclaim 5, wherein the biophotonic material is rigid.
 7. The biophotonicmaterial of claim 6, wherein the biophotonic material is substantiallytranslucent.
 8. The biophotonic material of claim 1, wherein thebiophotonic material has a translucency of at least about 40%, about50%, about 60%, about 70%, or about 80% at 460 nm.
 9. The biophotonicmaterial of claim 1, wherein the biophotonic material has a thickness ofabout 0.1 mm to about 50 mm.
 10. The biophotonic material of claim 1,wherein the biophotonic material has a pre-formed configuration.
 11. Thebiophotonic material of claim 10, wherein the pre-formed configurationis a shape and/or a size corresponding with a shape and/or a size of abody part to which the biophotonic material can be applied.
 12. Thebiophotonic material of claim 11, wherein the body part is selected froma head, scalp, forehead, nose, cheeks, ears, lip, face, neck, shoulder,arm pit, arm, elbow, hand, finger, abdomen, chest, stomach, back,sacrum, buttocks, genitals, legs, knee, feet, nails, hair, toes, boneyprominences, and combinations thereof.
 13. The biophotonic material ofclaim 1, wherein the biophotonic material is a mask.
 14. The biophotonicmaterial of claim 13, wherein the mask is a face mask having at leastone opening for the eyes, nose or mouth.
 15. The biophotonic material ofclaim 10, wherein the pre-formed configuration is a shape and/or a sizecorresponding with a shape and/or a size of a light source or lamp towhich the biophotonic material can be attached.
 16. The biophotonicmaterial of claim 1, wherein the biophotonic material can be removedwithout leaving substantially any residue on a surface to which thebiophotonic material is applied.
 17. The biophotonic material of claim1, wherein the at least one chromophore is a fluorophore.
 18. Thebiophotonic material of claim 17, wherein the fluorophore is a xanthenedye.
 19. The biophotonic material of claim 1, wherein the cohesivematrix comprises at least one polymer.
 20. The biophotonic material ofclaim 1, wherein the polymer is selected from a cross-linked polyacrylicpolymer, a hyaluronate, a hydrated polymer, a hydrophilic polymer. 21.The biophotonic material of claim 1, wherein the at least onechromophore is within the cohesive matrix.
 22. The biophotonic materialof claim 1, wherein the cohesive matrix is in particulate form.
 23. Abiophotonic material for phototherapy, said biophotonic materialcomprising at least one chromophore in a carrier medium, wherein thecarrier medium can form a cohesive matrix containing the at least onechromophore and wherein the at least one chromophore can absorb and emitlight within the cohesive matrix when illuminated with light.
 24. Thebiophotonic material of claim 23, wherein the carrier medium is at leastone polymer or a polymer pre-cursor which can form the cohesive matrixby polymerizing, cross-linking or drying. 25.-27. (canceled)
 28. Acontainer comprising: a sealed chamber for holding a biophotonicmaterial, and an outlet in communication with the chamber fordischarging the biophotonic material from the container, wherein thebiophotonic material comprises at least one chromophore in a carriermedium which can form a cohesive matrix after being discharged from thesealed chamber.
 29. A container according to claim 28, wherein thecontainer is a spray can.
 30. A kit comprising: a first componentcomprising a first chromophore; and a second component comprising athickening agent, wherein the thickening agent can form a cohesivematrix when mixed with the first component.
 31. A method for biophotonictreatment of a skin disorder comprising: placing a biophotonic materialover a target skin tissue, wherein the biophotonic material comprises atleast one chromophore and a cohesive matrix; and illuminating saidbiophotonic material with light having a wavelength that overlaps withan absorption spectrum of the at least one chromophore; wherein saidbiophotonic material emits fluorescence at a wavelength and intensitythat promotes healing of said skin disorder.
 32. A method forbiophotonic treatment of acne comprising: placing a biophotonic materialover a target skin tissue, wherein the biophotonic material comprises atleast one chromophore and a cohesive matrix; and illuminating saidbiophotonic material with light having a wavelength that overlaps withan absorption spectrum of the at least one chromophore; wherein saidbiophotonic material emits fluorescence at a wavelength and intensitythat treats the acne.
 33. A method for promoting wound healingcomprising: placing a biophotonic material over or within a wound,wherein the biophotonic material comprises at least one chromophore anda cohesive matrix; and illuminating said biophotonic material with lighthaving a wavelength that overlaps with an absorption spectrum of the atleast one chromophore; wherein said biophotonic material emitsfluorescence at a wavelength and intensity that promotes wound healing.34. A method for promoting skin rejuvenation comprising: placing abiophotonic material over a target skin tissue, wherein the biophotonicmaterial comprises at least one chromophore and a cohesive matrix; andilluminating said biophotonic material with light having a wavelengththat overlaps with an absorption spectrum of the at least onechromophore; wherein said biophotonic material emits fluorescence at awavelength and intensity that promotes skin rejuvenation. 35.-43.(canceled)